LOCUS       Exported                8256 bp ds-DNA     circular SYN 01-JUN-2015
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Cellecta-pRSI12N-U6-sh-HTS4-UbiC-TagRFP-2A-Neo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8256)
  AUTHORS   Peter
  TITLE     Direct Submission
  JOURNAL   Exported Jun 14, 2016 from SnapGene 2.7.3
            http://www.snapgene.com
COMMENT     This file is best viewed with a free Vector NTI Viewer:
            ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..8256
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        6..232
                     /note="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             233..413
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    460..585
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    1078..1311
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     promoter        1819..2059
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2065..2090
                     /note="cloning site"
                     /note="color: #ffff00"
     misc_feature    2138..2255
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     promoter        2336..2735
                     /note="UbiC promoter"
                     /note="human ubiquitin C promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2748..3458
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /note="TagRFP"
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             3465..3518
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2
                     Cleavage site after base 3515"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3525..4328
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #ccffcc"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4335..4923
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(4806..4817)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /note="color: #cc99b2
                     Cleavage site after base 4805"
                     /translation="IEGR"
     LTR             4997..5230
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    5302..5423
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      5463..5598
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     complement(5631..5647)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5655..5671
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(5679..5709)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5679 .. 5685 / #ffffff / -10
                        2: 5686 .. 5703 / #ffffff
                        3: 5704 .. 5709 / #ffffff / -35"
     protein_bind    5724..5745
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(6033..6621)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6792..7652)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6792 .. 7583 / #ccffcc
                        2: 7584 .. 7652 / #ccffcc / signal sequence
                     Cleavage site after base 7583"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7653..7757)
                     /gene="bla"
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     8231..8247
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 atggccgccc ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga
     1861 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
     1921 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     1981 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     2041 tatatcttgt ggaaaggacg aaacaccgga gtcttctttt ttgaagacac ttcggactgt
     2101 agaactctga acctctgaat tctcactcga gccacaattt taaaagaaaa ggggggattg
     2161 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     2221 aattacaaaa acaaattaca aaaattcaaa attttgcggt tgtgcttttt gcgtgtaaat
     2281 ctcggtggtc gccgtatcat ggcttgtgcg atgcgttggc taccggttct tcgaaggcct
     2341 ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc tcacggcgag cgctgccacg
     2401 tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2461 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2521 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2581 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2641 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2701 ttcttgtttg tggatcgctg tgatcgtcac ttggtctaga cgccaccatg gtgtctaagg
     2761 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     2821 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     2881 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     2941 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gacttcttta
     3001 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgag gacgggggcg
     3061 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     3121 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     3181 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga agcgacatgg
     3241 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     3301 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     3361 gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat
     3421 actgcgacct ccctagcaaa ctggggcaca aacttaatgg atccgagggc agaggaagcc
     3481 ttctaacatg cggtgacgtg gaggagaatc ccggcccttc cggaatggga tcggccattg
     3541 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3601 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3661 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3721 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3781 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3841 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3901 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3961 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4021 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgatg
     4081 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4141 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4201 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4261 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4321 tcttctgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     4381 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     4441 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     4501 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     4561 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     4621 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     4681 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     4741 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     4801 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4861 ctcttccgcg tctccgcctt cgccctcaga cgagtcggat ctctctttgg gccgcctccc
     4921 cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     4981 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga tctgcttttt
     5041 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5101 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5161 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5221 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     5281 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     5341 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5401 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     5461 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5521 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5581 gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat gtcatagctg
     5641 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5701 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5761 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5821 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7741 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     7801 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     7861 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     7921 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     7981 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     8041 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     8101 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     8161 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     8221 tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
