LOCUS V045733 10654 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045733 VERSION V045733 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 2815..5298 /label="GPAM(NM_001244949)" /note="GPAM(NM_001244949)" /gene="GPAM" CDS 5299..5325 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 5357..5943 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 5963..6562 /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 6576..7164 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7371..8004 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8132..8148) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8156..8172 /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(8180..8210) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8225..8246 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8534..9122) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9293..10153) /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(10154..10258) /gene="bla" /label="AmpR promoter" polyA_signal 10306..10440 /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 tctatttccg gtgaattcgc caccatggat 2821 gaatctgcac tgacccttgg tacaatagat gtttcttatc tgccacattc atcagaatac 2881 agtgttggtc gatgtaagca cacaagtgag gaatggggtg agtgtggctt tagacccacc 2941 atcttcagat ctgcaacttt aaaatggaaa gaaagcctaa tgagtcggaa aaggccattt 3001 gttggaagat gttgttactc ctgcactccc cagagctggg acaaattttt caaccccagt 3061 atcccgtctt tgggtttgcg gaatgttatt tatatcaatg aaactcacac aagacaccgc 3121 ggatggcttg caagacgcct ttcttacgtt ctttttattc aagagcgaga tgtgcataag 3181 ggcatgtttg ccaccaatgt gactgaaaat gtgctgaaca gcagtagagt acaagaggca 3241 attgcagaag tggctgctga attaaaccct gatggttctg cccagcagca atcaaaagcc 3301 gttaacaaag tgaaaaagaa agctaaaagg attcttcaag aaatggttgc cactgtctca 3361 ccggcaatga tcagactgac tgggtgggtg ctgctaaaac tgttcaacag cttcttttgg 3421 aacattcaaa ttcacaaagg tcaacttgag atggttaaag ctgcaactga gacgaatttg 3481 ccgcttctgt ttctaccagt tcatagatcc catattgact atctgctgct cactttcatt 3541 ctcttctgcc ataacatcaa agcaccatac attgcttcag gcaataatct caacatccca 3601 atcttcagta ccttgatcca taagcttggg ggcttcttca tacgacgaag gctcgatgaa 3661 acaccagatg gacggaaaga tgttctctat agagctttgc tccatgggca tatagttgaa 3721 ttacttcgac agcagcaatt cttggagatc ttcctggaag gcacacgttc taggagtgga 3781 aaaacctctt gtgctcgggc aggacttttg tcagttgtgg tagatactct gtctaccaat 3841 gtcatcccag acatcttgat aatacctgtt ggaatctcct atgatcgcat tatcgaaggt 3901 cactacaatg gtgaacaact gggcaaacct aagaagaatg agagcctgtg gagtgtagca 3961 agaggtgtta ttagaatgtt acgaaaaaac tatggttgtg tccgagtgga ttttgcacag 4021 ccattttcct taaaggaata tttagaaagc caaagtcaga aaccggtgtc tgctctactt 4081 tccctggagc aagcgttgtt accagctata cttccttcaa gacccagtga tgctgctgat 4141 gaaggtagag acacgtccat taatgagtcc agaaatgcaa cagatgaatc cctacgaagg 4201 aggttgattg caaatctggc tgagcatatt ctattcactg ctagcaagtc ctgtgccatt 4261 atgtccacac acattgtggc ttgcctgctc ctctacagac acaggcaggg aattgatctc 4321 tccacattgg tcgaagactt ctttgtgatg aaagaggaag tcctggctcg tgattttgac 4381 ctggggttct caggaaattc agaagatgta gtaatgcatg ccatacagct gctgggaaat 4441 tgtgtcacaa tcacccacac tagcaggaac gatgagtttt ttatcacccc cagcacaact 4501 gtcccatcag tcttcgaact caacttctac agcaatgggg tacttcatgt ctttatcatg 4561 gaggccatca tagcttgcag cctttatgca gttctgaaca agaggggact ggggggtccc 4621 actagcaccc cacctaacct gatcagccag gagcagctgg tgcggaaggc ggccagcctg 4681 tgctaccttc tctccaatga aggcaccatc tcactgcctt gccagacatt ttaccaagtc 4741 tgccatgaaa cagtaggaaa gtttatccag tatggcattc ttacagtggc agagcacgat 4801 gaccaggaag atatcagtcc tagtcttgct gagcagcagt gggacaagaa gcttccagaa 4861 cctttgtctt ggagaagtga tgaagaagat gaagacagtg actttgggga ggaacagcga 4921 gattgctacc tgaaggtgag ccaatccaag gagcaccagc agtttatcac cttcttacag 4981 agactccttg ggcctttgct ggaggcctac agctctgctg ccatctttgt tcacaacttc 5041 agtggtcctg ttccagaacc tgagtatctg caaaagttgc acaaatacct aataaccaga 5101 acagaaagaa atgttgcagt atatgctgag agtgccacat attgtcttgt gaagaatgct 5161 gtgaaaatgt ttaaggatat tggggttttc aaggagacca aacaaaagag agtgtctgtt 5221 ttagaactga gcagcacttt tctacctcaa tgcaaccgac aaaaacttct agaatatatt 5281 ctgagttttg tggtgctgta cccatacgat gttccagatt acgcttagac tagttctaga 5341 gcggccgcgg atcccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct 5401 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg 5461 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt 5521 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg 5581 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac 5641 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg 5701 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct 5761 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg 5821 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag 5881 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca 5941 acccacaagg agacgacctt ccatgaccga gtacaagccc acggtgcgcc tcgccacccg 6001 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 6061 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt 6121 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 6181 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 6241 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 6301 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga 6361 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 6421 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg 6481 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 6541 gacccgcaag cccggtgcct agacgcgtct ggaacaatca acctctggat tacaaaattt 6601 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 6661 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 6721 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 6781 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 6841 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 6901 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 6961 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc 7021 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 7081 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 7141 tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca 7201 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc 7261 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac 7321 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct 7381 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta 7441 cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg 7501 atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga 7561 gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt 7621 gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc 7681 ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt 7741 tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata 7801 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc 7861 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 7921 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 7981 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat 8041 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag cgtttaccgt 8101 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 8161 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 8221 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 8281 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 8341 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 8401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 8461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 8521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 8581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 8641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 8701 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 8761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 8821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 8881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 8941 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 9001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 9061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 9121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 9181 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 9241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 9301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 9361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 9421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 9481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 9541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 9601 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 9661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 9721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 9781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 9841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 9901 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 9961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 10021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 10081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 10141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 10201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 10261 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca 10321 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 10381 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc 10441 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc 10501 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca 10561 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt 10621 gtatttgtta aatatgtact acaaacttag tagt //