LOCUS V039432 8804 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039432 VERSION V039432 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..3560 /label="TIMP2(NM_003255)" /note="TIMP2(NM_003255)" /gene="TIMP2" promoter 3592..4091 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4112..4711 /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 4725..5313 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5520..6153 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6282..6298) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6306..6322 /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(6330..6360) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6375..6396 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6684..7272) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7443..8303) /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(8304..8408) /gene="bla" /label="AmpR promoter" polyA_signal 8456..8590 /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 atgggcgccg cggcccgcac cctgcggctg gcgctcggcc 2941 tcctgctgct ggcgacgctg cttcgcccgg ccgacgcctg cagctgctcc ccggtgcacc 3001 cgcaacaggc gttttgcaat gcagatgtag tgatcagggc caaagcggtc agtgagaagg 3061 aagtggactc tggaaacgac atttatggca accctatcaa gaggatccag tatgagatca 3121 agcagataaa gatgttcaaa gggcctgaga aggatataga gtttatctac acggccccct 3181 cctcggcagt gtgtggggtc tcgctggacg ttggaggaaa gaaggaatat ctcattgcag 3241 gaaaggccga gggggacggc aagatgcaca tcaccctctg tgacttcatc gtgccctggg 3301 acaccctgag caccacccag aagaagagcc tgaaccacag gtaccagatg ggctgcgagt 3361 gcaagatcac gcgctgcccc atgatcccgt gctacatctc ctccccggac gagtgcctct 3421 ggatggactg ggtcacagag aagaacatca acgggcacca ggccaagttc ttcgcctgca 3481 tcaagagaag tgacggctcc tgtgcgtggt accgcggcgc ggcgcccccc aagcaggagt 3541 ttctcgacat cgaggaccca taaggatccc gcgactctag ataattctac cgggtagggg 3601 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc 3661 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg 3721 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc 3781 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta 3841 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg 3901 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg 3961 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc 4021 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc 4081 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct 4141 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 4201 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 4261 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 4321 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 4381 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 4441 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 4501 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 4561 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 4621 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 4681 ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt 4741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 4801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 4861 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 4921 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 4981 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 5041 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 5101 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 5161 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 5221 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 5281 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct 5341 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg 5401 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag 5461 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact 5521 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac 5581 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact 5641 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa 5701 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga 5761 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga 5821 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc 5881 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc 5941 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag 6001 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 6061 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 6121 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc 6181 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc 6241 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 6301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 6481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 6541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 6721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 6781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 6841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 6901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 6961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 7141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 7501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 7561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 7741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 7801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 7861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 7921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 7981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 8041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 8221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8401 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 8461 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 8521 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 8581 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 8641 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 8701 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt 8761 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //