LOCUS Exported 8284 bp ds-DNA circular SYN 15-NOV-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 8284) AUTHORS Broad Institute TITLE Direct Submission JOURNAL Exported Friday, Nov 15, 2019 from SnapGene 5.0.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..8284 /organism="recombinant plasmid" /mol_type="other DNA" promoter 264..490 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 491..671 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 723..860 /label=psi /note="psi" misc_feature 827..1191 /label=gag /note="gag" misc_feature 1336..1569 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." promoter 1982..2222 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_feature 2209..2252 /label=Oligo_1210 misc_feature 2211..2231 /label=Argon misc_feature 2233..2252 /label=AsCpf1-DR misc_feature 2338..2455 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" misc_feature complement(2367..2391) /label=Kermit misc_feature complement(2571..2595) /label=Beaker promoter 2594..3772 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" CDS 3785..4384 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 4400..4988 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 5060..5293 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5371..5492 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 5532..5667 /label=SV40 ori /note="SV40 origin of replication" rep_origin 5874..6329 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 6355..6459 /gene="bla" /label=AmpR promoter CDS 6460..7320 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 7491..8079 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 181 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac 241 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat 301 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt 361 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg 421 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg 481 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 541 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 601 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 661 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc 721 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg 781 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc 841 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg 901 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc 961 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta 1021 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc 1081 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata 1141 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc 1201 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa 1261 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa 1321 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat 1381 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca 1441 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt 1501 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca 1561 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg 1621 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga 1681 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca 1741 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg 1801 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg 1861 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca 1921 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccagaggtac 1981 cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga 2041 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag 2101 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca 2161 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg 2221 acgaaacacc ggtaatttct actcttgtag atggagacgc gttttttctc gtctctgaat 2281 tcgctagcaa gcttggcgta actagatctt gagacaaatg gcagtattca tccacaattt 2341 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 2401 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt 2461 ttattacagg gacagcagag atccactttg gcgccggctc gagggggccc gggtgcaaag 2521 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa 2581 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc 2641 cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt 2701 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc 2761 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac 2821 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg 2881 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt 2941 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg 3001 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc 3061 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac 3121 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc 3181 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag 3241 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc 3301 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc 3361 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag 3421 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa 3481 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc 3541 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg 3601 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg 3661 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct 3721 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gacgtacggc 3781 caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc 3841 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc 3901 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 3961 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcc gtggcggtct ggaccacgcc 4021 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 4081 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 4141 ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct 4201 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 4261 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 4321 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 4381 ctgaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 4441 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 4501 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 4561 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 4621 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 4681 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 4741 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 4801 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 4861 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 4921 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 4981 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5041 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt 5101 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa 5161 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt 5221 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg 5281 aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt 5341 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac 5401 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 5461 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc 5521 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 5581 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 5641 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc 5701 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 5761 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 5821 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc 5881 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 5941 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 6001 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 6061 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 6121 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 6181 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 6241 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 6301 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga 6361 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 6421 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 6481 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 6541 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 6601 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 6661 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 6721 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 6781 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 6841 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 6901 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 6961 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 7021 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 7081 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 7141 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 7201 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 7261 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 7321 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 7381 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 7441 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 7501 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 7561 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 7621 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 7681 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 7741 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 7801 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 7861 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 7921 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 7981 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 8041 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 8101 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 8161 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 8221 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 8281 cctc //