LOCUS V035808 8827 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035808 VERSION V035808 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..3681 /label="CHRM1(NM_000738)" /note="CHRM1(NM_000738)" /gene="CHRM1" CDS 3688..3753 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3797..4008 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4021..4289 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4316..4915 /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 4922..5510 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5584..5817 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5889..6010 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6029..6164 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6202..6218) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6226..6242 /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(6250..6280) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6295..6316 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6604..7192) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7363..8223) /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(8224..8328) /gene="bla" /label="AmpR promoter" primer_bind 8802..8818 /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 catgaacact tcagccccac ctgctgtcag ccccaacatc 2341 accgtcctgg caccaggaaa gggtccctgg caagtggcct tcattgggat caccacgggc 2401 ctcctgtcgc tagccacagt gacaggcaac ctgctggtac tcatctcttt caaggtcaac 2461 acggagctca agacagtcaa taactacttc ctgctgagcc tggcctgtgc tgacctcatc 2521 atcggtacct tctccatgaa cctctatacc acgtacctgc tcatgggcca ctgggctctg 2581 ggcacgctgg cttgtgacct ctggctggcc ctggactatg tggccagcaa tgcctccgtc 2641 atgaatctgc tgctcatcag ctttgaccgc tacttctccg tgactcggcc cctgagctac 2701 cgtgccaagc gcacaccccg ccgggcagct ctgatgatcg gcctggcctg gctggtttcc 2761 tttgtgctct gggccccagc catcctcttc tggcagtacc tggtagggga gcggacagtg 2821 ctagctgggc agtgctacat ccagttcctc tcccagccca tcatcacctt tggcacagcc 2881 atggctgcct tctacctccc tgtcacagtc atgtgcacgc tctactggcg catctaccgg 2941 gagacagaga accgagcacg ggagctggca gcccttcagg gctccgagac gccaggcaaa 3001 gggggtggca gcagcagcag ctcagagagg tctcagccag gggctgaggg ctcaccagag 3061 actcctccag gccgctgctg tcgctgctgc cgggccccca ggctgctgca ggcctacagc 3121 tggaaggaag aagaggaaga ggacgaaggc tccatggagt ccctcacatc ctcagaggga 3181 gaggagcctg gctccgaagt ggtgatcaag atgccaatgg tggaccccga ggcacaggcc 3241 cccaccaagc agcccccacg gagctcccca aatacagtca agaggccgac taagaaaggg 3301 cgtgatcgag ctggcaaggg ccagaagccc cgtggaaagg agcagctggc caagcggaag 3361 accttctcgc tggtcaagga gaagaaggcg gctcggaccc tgagtgccat cctcctggcc 3421 ttcatcctca cctggacacc gtacaacatc atggtgctgg tgtccacctt ctgcaaggac 3481 tgtgttcccg agaccctgtg ggagctgggc tactggctgt gctacgtcaa cagcaccatc 3541 aaccccatgt gctacgcact ctgcaacaaa gccttccggg acacctttcg cctgctgctg 3601 ctttgccgct gggacaagag acgctggcgc aagatcccca agcgccctgg ctccgtgcac 3661 cgcactccct cccgccaatg cggatccgac tacaaagacc atgacggtga ttataaagat 3721 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 3781 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 3841 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 3901 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 3961 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 4021 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4081 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4141 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4201 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 4261 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 4321 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 4381 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 4441 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4501 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4561 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4621 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4681 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4741 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4801 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4861 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 4921 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4981 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5041 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5101 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5161 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5221 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5281 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 5341 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5401 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5461 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 5521 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5581 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5641 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5701 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5761 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5821 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 5881 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5941 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6001 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6061 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6121 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6181 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6301 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7021 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8341 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8401 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8461 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8521 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8581 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8641 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8701 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8761 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8821 caagctg //