LOCUS V035271 9244 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035271 VERSION V035271 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /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 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..4098 /label="Esr1(NM_007956)" /note="Esr1(NM_007956)" /gene="Esr1" CDS 4105..4170 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4214..4425 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4438..4706 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4733..5332 /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 5339..5927 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6001..6234 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6306..6427 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6446..6581 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6619..6635) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6643..6659 /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(6667..6697) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6712..6733 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7021..7609) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7780..8640) /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(8641..8745) /gene="bla" /label="AmpR promoter" primer_bind 9219..9235 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgaccatg acccttcaca ccaaagcctc gggaatggcc 2341 ttgctgcacc agatccaagg gaacgagctg gagcccctca accgcccgca gctcaagatg 2401 cccatggaga gggccctggg cgaggtatac gtggacaaca gcaagcccac tgtgttcaac 2461 taccccgagg gcgccgccta cgagttcaac gccgccgccg ccgccgccgc cgccgcctcg 2521 gcgccggtct acggccagtc gggcatcgcc tacggccccg ggtcggaggc ggccgccttc 2581 agtgccaaca gcctgggggc tttcccccag ctcaacagcg tgtcgcctag cccgctgatg 2641 ctgctgcacc cgccgccgca gctgtctcct ttcctgcacc cgcacggcca gcaggtgccc 2701 tactacctgg agaacgagcc cagcgcctac gccgtgcgcg acaccggccc tcccgccttc 2761 tacaggtcta attctgacaa tcgacgccag aatggccgag agagactgtc cagcagtaac 2821 gagaaaggaa acatgatcat ggagtctgcc aaggagactc gctactgtgc cgtgtgcaat 2881 gactatgcct ctggctacca ttatggggtc tggtcctgcg aaggctgcaa ggctttcttt 2941 aagagaagca ttcaaggaca caatgactac atgtgtccag ctacaaacca atgcaccatt 3001 gacaagaacc ggaggaagag ttgccaggcc tgtcggctgc gcaagtgtta cgaagtgggc 3061 atgatgaaag gcggcatacg gaaagaccgc cgaggaggga gaatgttgaa gcacaagcgt 3121 cagagagatg acttggaagg ccgaaatgaa atgggtgctt caggagacat gagggctgcc 3181 aacctttggc caagccctct tgtgattaag cacactaaga agaatagccc tgccttgtcc 3241 ttgacagctg accagatggt cagtgccttg ttggatgctg aaccgcccat gatctattct 3301 gaatatgatc cttctagacc cttcagtgaa gcctcaatga tgggcttatt gaccaaccta 3361 gcagataggg agctggttca tatgatcaac tgggcaaaga gagtgccagg ctttggggac 3421 ttgaatctcc atgatcaggt ccaccttctc gagtgtgcct ggctggagat tctgatgatt 3481 ggtctcgtct ggcgctccat ggaacacccg gggaagctcc tgtttgctcc taacttgctc 3541 ctggacagga atcaaggtaa atgtgtggaa ggcatggtgg agatctttga catgttgctg 3601 gctacgtcaa gtcggttccg catgatgaac ctgcagggag aagagtttgt gtgcctcaaa 3661 tccatcattt tgcttaattc cggagtgtac acgtttctgt ccagcacctt gaagtctctg 3721 gaagagaagg accacatcca ccgtgtcctg gacaagatca cagacacttt gatccacctg 3781 atggccaaag ctggcctgac tctgcagcag cagcatcgcc gcctagctca gctccttctc 3841 attctttccc atatccggca catgagtaac aaaggcatgg agcatctcta caacatgaaa 3901 tgcaagaacg ttgtgcccct ctatgacctg ctcctggaga tgttggatgc ccaccgcctt 3961 catgccccag ccagtcgcat gggagtgccc ccagaggagc ccagccagac ccagctggcc 4021 accaccagct ccacttcagc acattcctta caaacctact acataccccc ggaagcagag 4081 ggcttcccca acacgatcgg atccgactac aaagaccatg acggtgatta taaagatcat 4141 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 4201 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4261 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4321 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4381 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4441 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4501 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4561 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4621 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4681 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4741 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4801 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4861 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4921 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4981 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 5041 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 5101 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 5161 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5221 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5281 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5341 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5401 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5461 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5521 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5581 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5641 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5701 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5761 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5821 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5881 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5941 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 6001 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 6061 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6121 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6181 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6241 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6301 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6361 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6421 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6481 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6541 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6601 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6661 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6721 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6781 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7441 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8761 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8821 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8881 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8941 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9001 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9061 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9121 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9181 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9241 gctg //