LOCUS V037324 9607 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037324 VERSION V037324 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 2302..4461 /label="TGM5(NM_201631)" /note="TGM5(NM_201631)" /gene="TGM5" CDS 4468..4533 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4577..4788 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4801..5069 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5096..5695 /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 5702..6290 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6364..6597 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6669..6790 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6809..6944 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6982..6998) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7006..7022 /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(7030..7060) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7075..7096 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7384..7972) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8143..9003) /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(9004..9108) /gene="bla" /label="AmpR promoter" primer_bind 9582..9598 /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggcccaa gggctagaag tggccctcac agacctccag 2341 agctccagaa ataatgtgcg gcaccacacg gaggagatca ctgtggacca cctgcttgtt 2401 cgccggggcc aggccttcaa cctcaccctg tacttcagga accggagctt ccagccaggc 2461 ctggacaaca tcatcttcgt ggttgaaact ggaccgctgc cagacctggc cttggggact 2521 cgggctgtgt tcagcctggc acgccatcac agccccagcc cctggattgc ctggctggag 2581 accaatgggg ccacctccac agaggtgagc ttgtgcgctc ctcccacggc ggccgtgggt 2641 cggtacctct tgaaaatcca catcgactcc ttccaggggt ctgtgacggc ctaccagcta 2701 ggggagttca tcctgctttt caatccctgg tgcccagagg atgctgtcta cttggacagt 2761 gaaccccaga ggcaggagta tgtcatgaat gattatggct tcatctacca aggcagcaag 2821 aactggatcc gcccatgtcc ctggaactat ggacagtttg aagacaaaat catagacatc 2881 tgcctgaagc tgctagacaa gagcctgcac ttccagactg acccagccac agactgtgct 2941 ctgcggggaa gccccgtcta cgtcagcaga gtggtgtgtg ccatgatcaa cagcaatgat 3001 gataatgggg tgctcaatgg aaactggagt gagaattaca cagacggcgc caaccctgcg 3061 gagtggacgg gcagcgtggc catcctgaag cagtggaacg ccacaggctg ccagcccgtg 3121 cgctacgggc aatgctgggt ctttgctgcc gtcatgtgca cagtgatgag gtgtctgggg 3181 atccctaccc gtgtgatcac caacttcgac tctggccacg atacagatgg aaacctgatc 3241 atagatgagt attatgacaa cacaggcagg attttgggga ataagaagaa ggatactatc 3301 tggaacttcc atgtctggaa tgagtgctgg atggcccgga aggatctgcc ccctgcatat 3361 ggaggctggc aggtgctgga cgccacacct caggagatga gcaacggcgt ctactgctgt 3421 ggccctgcct ctgtcagagc catcaaagaa ggagaagtgg acctgaacta tgacacgccc 3481 tttgtgtttt cgatggtgaa tgctgactgc atgtcctggc tcgtccaggg agggaaggag 3541 cagaagcttc accaggacac gagttctgtt ggcaatttta tcagcacaaa gagcatccag 3601 agtgacgagc gggatgacat cacagagaac tacaagtatg aagaaggatc cctccaggag 3661 aggcaggtgt ttctgaaggc tctgcagaag ctgaaggcta gaagcttcca tggctcccaa 3721 agaggagcag agttgcaacc ttccaggccc acatcactga gccaggacag ccctcggagc 3781 ctgcatacac cttcccttcg acccagtgat gtggtgcaag tctccctgaa attcaagctg 3841 ctcgacccgc ccaacatggg ccaggatata tgctttgtcc tgctggccct caacatgtcc 3901 tcccagttca aggacctcaa agtgaacctg agtgcccagt ctctgctgca cgatggcagc 3961 cccctgtccc cattctggca ggacacagcg ttcatcacac tctctcctaa agaagcaaag 4021 acctacccct gcaaaatctc ctattcccag tacagccagt acctgtcaac agacaagctg 4081 atccgcatca gtgccctggg tgaagagaaa agcagtcctg agaaaatcct ggtgaacaag 4141 atcatcacct tatcttatcc aagcatcacg attaatgttc taggagcagc cgttgtgaac 4201 cagccactct ccatacaggt gatattttca aaccccctct cggagcaggt tgaggactgt 4261 gtgctgactg tggaaggaag tggcctcttc aagaaacagc agaaagtctt ccttggagtc 4321 ctcaaacccc aacaccaagc aagcatcatt ctggagaccg tccccttcaa gagtggacaa 4381 aggcagatcc aagctaatat gagaagcaac aagtttaagg acattaaggg ttacaggaat 4441 gtttatgtag actttgcatt aggatccgac tacaaagacc atgacggtga ttataaagat 4501 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 4561 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 4621 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 4681 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 4741 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 4801 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4861 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4921 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4981 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 5041 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 5101 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 5161 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 5221 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 5281 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 5341 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 5401 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 5461 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 5521 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 5581 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 5641 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 5701 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5761 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5821 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5881 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5941 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 6001 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 6061 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 6121 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 6181 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 6241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 6301 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 6361 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 6421 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6481 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6541 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6601 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6661 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6721 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6781 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6841 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6901 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6961 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 7021 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7081 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7141 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7201 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7261 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7321 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7381 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7441 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7501 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7561 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7621 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7681 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7741 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7801 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7861 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7921 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7981 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8041 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8281 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9001 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9061 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9121 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9181 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9241 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9301 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9361 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9421 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9481 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9541 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9601 caagctg //