LOCUS V022739 8556 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022739 VERSION V022739 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2301..3410 /label="USP12(NM_182488)" /note="USP12(NM_182488)" /gene="USP12" CDS 3417..3482 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3526..3737 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3750..4018 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4045..4644 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" misc_feature 4651..5239 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5313..5546 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5618..5739 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5758..5893 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5931..5947) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5955..5971 /bound_moiety="lac repressor encoded by lacI" /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(5979..6009) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6024..6045 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6333..6921) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7092..7952) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(7953..8057) /gene="bla" /label="AmpR promoter" primer_bind 8531..8547 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" 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 cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcgccacc atggaaatcc taatgacagt ctccaaattc gcctccatct 2341 gtaccatggg cgccaatgct tcggcattag agaaagagat tggtccagaa cagtttccgg 2401 tcaatgagca ctattttgga ttagtcaatt ttgggaatac ctgctactgc aattcagttc 2461 ttcaagcact ttatttttgt cgtccatttc gggaaaaagt tcttgcgtat aagagtcaac 2521 ctaggaaaaa ggagagcctt cttacatgct tagcagatct cttccatagc atagccactc 2581 agaagaaaaa ggttggagta atacccccta agaagttcat cacaagatta cggaaagaaa 2641 atgagctttt tgacaactac atgcaacaag atgcccatga attcttaaat tacctactaa 2701 atacaattgc tgatatttta caagaagaga gaaagcagga aaaacaaaat ggtcgtttac 2761 ctaatggtaa tattgataat gaaaataata acagcacacc agacccaacg tgggttcatg 2821 agatttttca gggaacatta actaatgaaa ccagatgtct tacttgtgaa actataagca 2881 gcaaagatga agatttttta gacctttctg ttgacgtgga acaaaataca tcaattactc 2941 actgcttaag gggtttcagc aacacagaaa ctctgtgcag tgaatacaag tattactgtg 3001 aagagtgtcg cagcaaacag gaagcacaca aacggatgaa agttaaaaaa ctgcccatga 3061 ttctagctct acacctgaag agatttaaat atatggatca acttcatcga tatacaaaac 3121 tctcttaccg ggtagttttt cctttagaac ttcgtctgtt taacacttca ggtgatgcca 3181 ccaatccaga cagaatgtac gaccttgttg ctgttgtggt tcactgtgga agtggtccca 3241 atcgaggcca ttatattgca atagttaaga gtcatgattt ttggttgttg tttgatgacg 3301 acattgtaga aaaaatagat gcacaagcta ttgaagaatt ctacgggttg acatcagata 3361 tctcaaagaa ctctgagtct ggttacatcc ttttctatca gtctcgggac ggatccgact 3421 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca 3481 agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat 3541 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa 3601 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg 3661 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt 3721 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc 3781 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg 3841 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc 3901 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga 3961 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca 4021 agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc 4081 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 4141 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 4201 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 4261 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 4321 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 4381 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 4441 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 4501 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 4561 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 4621 atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa 4681 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 4741 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 4801 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 4861 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 4921 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 4981 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 5041 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 5101 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 5161 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 5221 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga 5281 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa 5341 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga 5401 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 5461 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 5521 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt 5581 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa 5641 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 5701 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc 5761 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 5821 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 5881 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg 5941 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 6001 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 6061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 6121 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 6181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 6241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 6301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 6361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 6421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 6481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 6541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 6601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 6661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 6721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 6781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 6901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 6961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 7021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 7081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 7141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 7201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 7261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 7321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 7381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 7441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 7501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 7561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 7621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 7681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 7741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 7801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 7861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 7921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 7981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 8041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt 8101 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt 8161 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt 8221 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg 8281 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg 8341 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca 8401 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 8461 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 8521 tcacgacgtt gtaaaacgac ggccagtgcc aagctg //