LOCUS       Exported                7248 bp ds-DNA     circular SYN 15-JUN-2016
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Cellecta-pRSIUP-U6-sh-HTS6-UbiC-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7248)
  AUTHORS   Cellecta
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2016 from SnapGene 2.7.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7248
                     /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..2064
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2065..2094
                     /note="Cloning Site"
                     /note="color: #ffff00"
     misc_feature    2122..2237
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
                     /note="color: #ffe4c4"
     promoter        2303..2702
                     /note="UbiC promoter"
                     /note="human ubiquitin C promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2715..3314
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    3327..3915
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(3798..3809)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /note="color: #cc99b2
                     Cleavage site after base 3797"
                     /translation="IEGR"
     LTR             3989..4222
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    4294..4415
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      4455..4590
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     complement(4623..4639)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4647..4663
                     /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(4671..4701)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 4671 .. 4677 / #ffffff / -10
                        2: 4678 .. 4695 / #ffffff
                        3: 4696 .. 4701 / #ffffff / -35"
     protein_bind    4716..4737
                     /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(5025..5613)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5784..6644)
                     /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: 5784 .. 6575 / #ccffcc
                        2: 6576 .. 6644 / #ccffcc / signal sequence
                     Cleavage site after base 6575"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6645..6749)
                     /gene="bla"
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     7223..7239
                     /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 acctcgagca atttaaaaga aaagggggga ttggggggta cagtgcaggg
     2161 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2221 acaaaaattc aaaatttctg cgttgttgtc ggtgctcgtt ctctgctctt cacgctactg
     2281 aattcatcac cggttcttcg aaggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     2341 cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg tcctgatcct
     2401 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     2461 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     2521 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     2581 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     2641 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     2701 gtctagacgc caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     2761 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     2821 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     2881 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     2941 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     3001 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     3061 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     3121 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     3181 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     3241 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     3301 agcccggtgc ctgaatctag gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     3361 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3421 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3481 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3541 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3601 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3661 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3721 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3781 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3841 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctctcttt
     3901 gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     3961 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagataa
     4021 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     4081 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     4141 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     4201 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     4261 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     4321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4381 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     4441 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4501 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4561 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa attcgtaatc
     4621 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4681 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4741 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4801 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5461 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5761 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5821 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5881 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5941 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6001 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6061 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6121 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6181 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6241 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6301 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6361 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6421 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6481 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6541 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6601 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6661 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6721 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6781 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6841 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6901 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6961 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     7021 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     7081 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7141 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7201 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
