LOCUS V039443 10124 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039443 VERSION V039443 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" regulatory 2820..2829 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2829..2894 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2901..4880 /label="MMP2(NM_004530)" /note="MMP2(NM_004530)" /gene="MMP2" promoter 4912..5411 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5432..6031 /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 6045..6633 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6840..7473 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7602..7618) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7626..7642 /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(7650..7680) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7695..7716 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8004..8592) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8763..9623) /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(9624..9728) /gene="bla" /label="AmpR promoter" polyA_signal 9776..9910 /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 ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg 2881 atgacgatga caaggaattc atggaggcgc taatggcccg gggcgcgctc acgggtcccc 2941 tgagggcgct ctgtctcctg ggctgcctgc tgagccacgc cgccgccgcg ccgtcgccca 3001 tcatcaagtt ccccggcgat gtcgccccca aaacggacaa agagttggca gtgcaatacc 3061 tgaacacctt ctatggctgc cccaaggaga gctgcaacct gtttgtgctg aaggacacac 3121 taaagaagat gcagaagttc tttggactgc cccagacagg tgatcttgac cagaatacca 3181 tcgagaccat gcggaagcca cgctgcggca acccagatgt ggccaactac aacttcttcc 3241 ctcgcaagcc caagtgggac aagaaccaga tcacatacag gatcattggc tacacacctg 3301 atctggaccc agagacagtg gatgatgcct ttgctcgtgc cttccaagtc tggagcgatg 3361 tgaccccact gcggttttct cgaatccatg atggagaggc agacatcatg atcaactttg 3421 gccgctggga gcatggcgat ggatacccct ttgacggtaa ggacggactc ctggctcatg 3481 ccttcgcccc aggcactggt gttgggggag actcccattt tgatgacgat gagctatgga 3541 ccttgggaga aggccaagtg gtccgtgtga agtatgggaa cgccgatggg gagtactgca 3601 agttcccctt cttgttcaat ggcaaggagt acaacagctg cactgatacc ggccgcagcg 3661 atggcttcct ctggtgctcc accacctaca actttgagaa ggatggcaag tacggcttct 3721 gtccccatga agccctgttc accatgggcg gcaacgctga aggacagccc tgcaagtttc 3781 cattccgctt ccagggcaca tcctatgaca gctgcaccac tgagggccgc acggatggct 3841 accgctggtg cggcaccact gaggactacg accgcgacaa gaagtatggc ttctgccctg 3901 agaccgccat gtccactgtt ggtgggaact cagaaggtgc cccctgtgtc ttccccttca 3961 ctttcctggg caacaaatat gagagctgca ccagcgccgg ccgcagtgac ggaaagatgt 4021 ggtgtgcgac cacagccaac tacgatgatg accgcaagtg gggcttctgc cctgaccaag 4081 ggtacagcct gttcctcgtg gcagcccacg agtttggcca cgccatgggg ctggagcact 4141 cccaagaccc tggggccctg atggcaccca tttacaccta caccaagaac ttccgtctgt 4201 cccaggatga catcaagggc attcaggagc tctatggggc ctctcctgac attgaccttg 4261 gcaccggccc cacccccacg ctgggccctg tcactcctga gatctgcaaa caggacattg 4321 tatttgatgg catcgctcag atccgtggtg agatcttctt cttcaaggac cggttcattt 4381 ggcggactgt gacgccacgt gacaagccca tggggcccct gctggtggcc acattctggc 4441 ctgagctccc ggaaaagatt gatgcggtat acgaggcccc acaggaggag aaggctgtgt 4501 tctttgcagg gaatgaatac tggatctact cagccagcac cctggagcga gggtacccca 4561 agccactgac cagcctggga ctgccccctg atgtccagcg agtggatgcc gcctttaact 4621 ggagcaaaaa caagaagaca tacatctttg ctggagacaa attctggaga tacaatgagg 4681 tgaagaagaa aatggatcct ggcttcccca agctcatcgc agatgcctgg aatgccatcc 4741 ccgataacct ggatgccgtc gtggacctgc agggcggcgg tcacagctac ttcttcaagg 4801 gtgcctatta cctgaagctg gagaaccaaa gtctgaagag cgtgaagttt ggaagcatca 4861 aatccgactg gctaggctgc tgaggatccc gcgactctag ataattctac cgggtagggg 4921 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc 4981 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg 5041 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc 5101 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta 5161 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg 5221 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg 5281 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc 5341 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc 5401 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct 5461 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 5521 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 5581 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 5641 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 5701 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 5761 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 5821 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 5881 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 5941 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 6001 ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt 6061 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 6121 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 6181 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 6241 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 6301 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 6361 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 6421 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 6481 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 6541 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 6601 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct 6661 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg 6721 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag 6781 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact 6841 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac 6901 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact 6961 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa 7021 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga 7081 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga 7141 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc 7201 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc 7261 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag 7321 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 7381 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 7441 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc 7501 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc 7561 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 7621 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 7681 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 7741 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 7801 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 7861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 7921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 7981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 8041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 8101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 8161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 8221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 8281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 8341 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 8401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 8461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 8521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 8581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 8641 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 8701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 8761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 8821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 8881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 8941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 9001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 9061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 9121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 9181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 9241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 9301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 9361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 9421 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 9481 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 9541 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 9601 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 9661 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 9721 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 9781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 9841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 9901 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 9961 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 10021 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt 10081 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //