LOCUS       Exported                7963 bp ds-DNA     circular SYN 15-JUN-2016
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Cellecta-pRSI20-U6-sh-HTS6-CMV-TagRFP-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7963)
  AUTHORS   Cellecta
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2016 from SnapGene 2.7.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7963
                     /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"
                     /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"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
                     /note="color: #ffe4c4"
     promoter        2295..2638
                     /note="CMV"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2653..3363
                     /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             3370..3423
                     /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 3420"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3430..4029
                     /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    4042..4630
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(4513..4524)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /note="color: #cc99b2
                     Cleavage site after base 4512"
                     /translation="IEGR"
     LTR             4704..4937
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    5009..5130
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      5170..5305
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     complement(5338..5354)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5362..5378
                     /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(5386..5416)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5386 .. 5392 / #ffffff / -10
                        2: 5393 .. 5410 / #ffffff
                        3: 5411 .. 5416 / #ffffff / -35"
     protein_bind    5431..5452
                     /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(5740..6328)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6499..7359)
                     /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: 6499 .. 7290 / #ccffcc
                        2: 7291 .. 7359 / #ccffcc / signal sequence
                     Cleavage site after base 7290"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7360..7464)
                     /gene="bla"
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     7938..7954
                     /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 cggtattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2341 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2401 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgccaatgg
     2461 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2521 attgacgcaa atgggcggta ggcgtgtacg gtggaaggtt tatataagca gagctcgttt
     2581 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagatt
     2641 ctagacgcca ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag
     2701 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
     2761 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
     2821 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac
     2881 acccagggca tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga
     2941 gtcaccacat acgaggacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     3001 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     3061 atgcagaaga aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc
     3121 ggcctggaag gcagaagcga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     3181 aacttcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     3241 tactatgtgg accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     3301 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt
     3361 aatggatccg agggcagagg aagccttcta acatgcggtg acgtggagga gaatcccggc
     3421 ccttccggga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3481 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3541 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3601 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3661 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3721 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     3781 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3841 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     3901 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3961 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4021 ggtgcctgaa tctaggtcga caatcaacct ctggattaca aaatttgtga aagattgact
     4081 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4141 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4201 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4261 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4321 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4381 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4441 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4501 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4561 ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc tctttgggcc
     4621 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     4681 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct
     4741 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4801 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4861 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4921 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     4981 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     5161 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5221 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5281 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc
     5341 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5401 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5461 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5521 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6181 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7381 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7441 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     7501 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     7561 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     7621 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     7681 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     7741 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     7801 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     7861 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     7921 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
