LOCUS V032603 9874 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032603 VERSION V032603 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 2843..4570 /label="Nt5e(NM_011851)" /note="Nt5e(NM_011851)" /gene="Nt5e" CDS 4577..4642 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4662..5161 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5182..5781 /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 5795..6383 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6590..7223 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7352..7368) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7376..7392 /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(7400..7430) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7445..7466 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7754..8342) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8513..9373) /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(9374..9478) /gene="bla" /label="AmpR promoter" polyA_signal 9526..9660 /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 ctcaagcttc gaattcgcca ccatgcgtcc cgcggccgct aaggtaccca agtggctgct 2881 tctcgcactg agcgctctac taccacagtg gcccgcagcc agtgcctggg agctcacgat 2941 cctgcacaca aacgacgtgc acagccggct agagcagacc agcgatgact ccaccaagtg 3001 cctcaacgcc agcctgtgtg tgggcggcgt ggcccggctc tttaccaagg tgcagcagat 3061 ccgcaaggaa gaacccaacg tgctgttttt ggatgccgga gaccagtacc agggcaccat 3121 ctggttcacc gtttacaaag gccttgaagt ggcacacttc atgaacatcc tgggctacga 3181 tgctatggca ctgggaaatc atgaatttga taacggtgtg gaaggactga ttgatcccct 3241 cctcagaaac gttaaatttc caattctgag cgcaaacatt aaggcacggg ggcctctagc 3301 acatcagata tctggacttt ttctgccatc taaagttctc tctgttggcg gtgaggttgt 3361 ggggattgtt ggatatactt caaaggaaac ccctttcctc tcaaatccag ggacaaattt 3421 agtctttgaa gatgaaatct ctgcattgca gcctgaagta gataaactaa agacgctaaa 3481 tgtgaataag atcatcgccc tggggcactc tggttttgag atggacaaac ttatcgctca 3541 gaaagttcga ggtgtggaca tcgtggtggg aggacactcc aacacctttc tctacacagg 3601 aaatccacct tccaaagaag tgcctgcggg gaagtaccca ttcatagtca ccgcagatga 3661 tggacggcag gtgcctgtgg tccaggccta tgcctttggc aaatacctgg gctacctgaa 3721 ggttgagttt gatgataaag gcaatgttat cacttcctat ggaaatccca ttcttctcaa 3781 cagcagcatt cctgaagatg cgaccatcaa agcagacatt aaccaatgga ggataaaatt 3841 agataattat tctacccagg aactcgggag aacgatcgtc tacctggatg gctccactca 3901 gacgtgccgc ttcagggaat gcaacatggg aaacctgatc tgtgatgcca tgattaacaa 3961 caacctcaga cacccagatg aaatgttttg gaaccacgtg tccatgtgca ttgtaaacgg 4021 aggtggcatc cggtccccca ttgatgagaa gaacaatggt accatcacct gggagaacct 4081 ggctgctgtg ctgccctttg gagggacatt tgacctcgtc caattaaaag ggtccaccct 4141 gaagaaggct tttgagcaca gcgtgcatcg ctatggccag tccaccggag agttcctgca 4201 agtgggtgga atccatgtgg tgtacgatat taaccgaaag ccctggaaca gagtggtcca 4261 attagaagtt ctctgcacca agtgtcgagt gcccatctat gagcctcttg aaatggataa 4321 agtgtataaa gtgaccctcc caagctatct ggccaacggt ggagatggat tccagatgat 4381 aaaagatgaa ttactaaagc atgactctgg tgatcaagat atcagcgtgg tttctgaata 4441 catctcaaaa atgaaagtag tttacccagc cgttgaaggg cggatcaagt tctctgcagc 4501 aagtcattac cagggaagct ttcctttagt aattctttca ttttgggcaa tgatccttat 4561 tttgtaccaa ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga 4621 ctacaaggat gacgatgaca agtagtctag ataattctac cgggtagggg aggcgctttt 4681 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag 4741 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt 4801 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc 4861 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca 4921 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca 4981 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct 5041 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg 5101 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc 5161 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc 5221 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg 5281 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc 5341 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg 5401 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg 5461 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg 5521 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac 5581 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc 5641 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg 5701 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg 5761 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg 5821 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 5881 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 5941 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 6001 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 6061 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 6121 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 6181 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 6241 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 6301 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 6361 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat 6421 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca 6481 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact 6541 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta 6601 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac 6661 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga 6721 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag 6781 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg 6841 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg 6901 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt 6961 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa 7021 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct 7081 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 7141 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 7201 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata 7261 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt 7321 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 7381 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 7441 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7501 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 7561 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 7741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 7801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 7861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 7921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 7981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 8041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 8101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 8161 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 8221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 8281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 8341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 8401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 8461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 8521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 8581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 8701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 8761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 8821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 8881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 8941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 9001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 9061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 9121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 9181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 9241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 9301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 9361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 9421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 9481 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca 9541 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 9601 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc 9661 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc 9721 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca 9781 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt 9841 gtatttgtta aatatgtact acaaacttag tagt //