LOCUS V023905 10868 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023905 VERSION V023905 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2826..5594 /label="CCAR2(NM_021174)" /note="CCAR2(NM_021174)" /gene="CCAR2" CDS 5601..5624 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 5656..6155 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6176..6775 /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 6789..7377 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7584..8217 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8346..8362) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8370..8386 /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(8394..8424) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8439..8460 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8748..9336) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9507..10367) /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(10368..10472) /gene="bla" /label="AmpR promoter" polyA_signal 10520..10654 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ccaccatgtc ccagtttaag cgccagcgga tcaacccgct tccaggggga cgcaacttct 2881 caggcacagc ttcaacatct cttctgggcc ctcctcctgg tttgctcact cctcctgtgg 2941 ccacagaact gtcccagaat gccaggcacc ttcagggtgg ggagaaacag cgggtcttca 3001 ctggtattgt taccagcttg catgactact ttggggttgt ggatgaagag gtcttttttc 3061 agctaagtgt ggtgaagggc cgtctgcccc agctgggtga gaaggtgctg gtgaaggctg 3121 catacaaccc aggccaggca gtgccctgga atgctgtcaa ggtgcaaacg ctctccaacc 3181 agcccctact gaagtcccca gcacctcctc ttctgcatgt agcagccctg ggccagaagc 3241 aagggatcct gggagctcag cctcagttga tcttccagcc tcaccggatt cccccactct 3301 ttcctcagaa gcctctgagt ctcttccaaa catcccacac acttcacctg agccacctga 3361 acagatttcc tgcccggggc cctcatggac ggttggatca gggccgaagt gatgactatg 3421 actccaagaa acgcaaacag cgggctggtg gagagccctg gggtgctaag aagccaaggc 3481 atgacctgcc tccttaccgg gtccacctca ctccttacac tgtggacagc cccatctgtg 3541 acttcctaga actccagcgc cgttaccgca gcctcctggt cccctcagat tttctgtccg 3601 tgcatctgag ttggctatca gccttccccc tgagccagcc cttttccctc catcatccaa 3661 gccggatcca ggtctcttct gaaaaggagg cagctccaga cgctggtgct gagcccatca 3721 ctgcagacag tgaccccgct tatagttcga aggtactgct gctctcttcc ccggggttgg 3781 aggaattgta tcgttgttgc atgctctttg tggatgacat ggctgagcca agggagacgc 3841 cagagcatcc tctgaagcag attaagtttt tgctgggcag gaaagaagag gaggcagtgc 3901 tggttggggg tgaatggtct ccttccctgg atggcctcga cccccaggct gacccgcagg 3961 tgctggtgcg taccgccatc cgctgtgcgc aggcccagac tggcattgat ttgagcggct 4021 gtaccaagtg gtggcgcttt gccgagtttc agtacctgca gccgggaccc ccccggcggc 4081 ttcagacagt ggtggtgtac ctgccggatg tctggaccat catgcctact ttggaggagt 4141 gggaggccct gtgccagcag aaagctgcag aggcagctcc cccaacccag gaggcacaag 4201 gggaaacgga gcctactgaa caggcacctg atgccttgga gcaagcagca gacacttcta 4261 gacggaacgc agaaactcca gaggccacca cacagcagga aacggacact gatctcccag 4321 aggcccctcc acccccccta gaacctgctg tcatcgcacg ccctggctgt gtaaacctgt 4381 ccctccatgg gattgtggag gatcggaggc caaaggaaag gatctctttt gaggtgatgg 4441 tgctggccga gctgtttctg gagatgctcc agagggattt tggctataga gtttataaga 4501 tgctactgag ccttcctgaa aaggtcgtgt ccccacctga acctgagaag gaggaggcgg 4561 ccaaggaaga agccaccaag gaggaagaag ccatcaaaga ggaggtggtc aaggagccca 4621 aggatgaggc acagaatgag ggcccggcta cagagtcaga ggccccgctg aaggaggatg 4681 ggcttttgcc caaaccactc tcttctgggg gagaggaaga agaaaaaccc cggggcgagg 4741 cttctgagga cctgtgtgag atggccctgg acccagaact gttgcttctg agggatgatg 4801 gagaggagga gtttgcagga gcaaagctgg aggattcgga ggtccggtcc gttgcctcaa 4861 accagtcaga gatggagttc tcttcacttc aggacatgcc caaggagctg gatccctctg 4921 ctgtgctccc cttagactgt ctgcttgctt ttgtgttctt tgatgccaac tggtgtggct 4981 acttgcaccg gcgagactta gagaggatcc tccttaccct tgggatccgg ctcagtgcag 5041 agcaggccaa gcagctggtc agcagggtgg tgacccagaa catctgccag taccggagcc 5101 ttcagtacag ccgccaggag ggcctggatg gtggccttcc cgaggaggtg ctcttcggaa 5161 acctggacct gctgccccct cctgggaaaa gcacgaagcc aggtgctgcc cccacagaac 5221 acaaagcctt ggtgtcccac aatggcagcc tgattaacgt ggggagcctg ctgcagcgcg 5281 cggagcagca ggacagcggc cggctctacc tagagaacaa gatccacaca ctggagctga 5341 agctggagga gagccataac cgtttctcag ccactgaagt aaccaataag acgctggcgg 5401 cagagatgca ggagctgcga gtccggctgg cggaggccga ggagaccgcc cggacggcgg 5461 agcgacagaa gagccagctc cagcggctgc tgcaggagct ccgcaggcgt ctgacccccc 5521 tgcagctgga gatccagcgg gtggtggaaa aggctgacag ctgggtggag aaggaggagc 5581 cggcacctag caacgaattc gattacaagg atgacgacga taagtgagga tcccgcgact 5641 ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt 5701 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca 5761 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta 5821 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac 5881 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga 5941 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc 6001 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg 6061 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg 6121 cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac 6181 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac 6241 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca 6301 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 6361 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 6421 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 6481 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 6541 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 6601 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 6661 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 6721 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaccgcg 6781 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 6841 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 6901 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 6961 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 7021 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 7081 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 7141 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 7201 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 7261 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 7321 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 7381 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc 7441 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc 7501 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 7561 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat 7621 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc 7681 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga 7741 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag 7801 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct 7861 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg 7921 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac 7981 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt 8041 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 8101 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 8161 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 8221 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 8281 gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag cttggcgtaa 8341 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 8401 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 8461 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 8521 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 8581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 8641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 8701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 8761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 8821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 8881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 8941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 9001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 9061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 9121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 9181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 9241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 9301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 9361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 9421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 9481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 9541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 9601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 9661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 9721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 9781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 9841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 9901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 9961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 10021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 10081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 10141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 10201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 10261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 10321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 10381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 10441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 10501 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa 10561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 10621 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa 10681 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt 10741 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat 10801 atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg tactacaaac 10861 ttagtagt //