LOCUS V045188 9497 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045188 VERSION V045188 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" regulatory 2288..2297 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2297..2362 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2369..3493 /label="GPER1(NM_001505)" /note="GPER1(NM_001505)" /gene="GPER1" CDS 3506..4225 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" promoter 4266..4477 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4490..4758 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4791..5585 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5592..6180 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6254..6487 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6559..6680 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6699..6834 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6872..6888) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6896..6912 /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(6920..6950) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6965..6986 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7274..7862) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8033..8893) /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(8894..8998) /gene="bla" /label="AmpR promoter" primer_bind 9472..9488 /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2341 ctacaaggat gacgatgaca aggaattcat ggatgtgact tcccaagccc ggggcgtggg 2401 cctggagatg tacccaggca ccgcgcagcc tgcggccccc aacaccacct cccccgagct 2461 caacctgtcc cacccgctcc tgggcaccgc cctggccaat gggacaggtg agctctcgga 2521 gcaccagcag tacgtgatcg gcctgttcct ctcgtgcctc tacaccatct tcctcttccc 2581 catcggcttt gtgggcaaca tcctgatcct ggtggtgaac atcagcttcc gcgagaagat 2641 gaccatcccc gacctgtact tcatcaacct ggcggtggcg gacctcatcc tggtggccga 2701 ctccctcatt gaggtgttca acctgcacga gcggtactac gacatcgccg tcctgtgcac 2761 cttcatgtcg ctcttcctgc aggtcaacat gtacagcagc gtcttcttcc tcacctggat 2821 gagcttcgac cgctacatcg ccctggccag ggccatgcgc tgcagcctgt tccgcaccaa 2881 gcaccacgcc cggctgagct gtggcctcat ctggatggca tccgtgtcag ccacgctggt 2941 gcccttcacc gccgtgcacc tgcagcacac cgacgaggcc tgcttctgtt tcgcggatgt 3001 ccgggaggtg cagtggctcg aggtcacgct gggcttcatc gtgcccttcg ccatcatcgg 3061 cctgtgctac tccctcattg tccgggtgct ggtcagggcg caccggcacc gtgggctgcg 3121 gccccggcgg cagaaggcgc tccgcatgat cctcgcggtg gtgctggtct tcttcgtctg 3181 ctggctgccg gagaacgtct tcatcagcgt gcacctcctg cagcggacgc agcctggggc 3241 cgctccctgc aagcagtctt tccgccatgc ccaccccctc acgggccaca ttgtcaacct 3301 cgccgccttc tccaacagct gcctaaaccc cctcatctac agctttctcg gggagacctt 3361 cagggacaag ctgaggctgt acattgagca gaaaacaaat ttgccggccc tgaaccgctt 3421 ctgtcacgct gccctgaagg ccgtcattcc agacagcacc gagcagtcgg atgtgaggtt 3481 cagcagtgcc gtgggatccg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 3541 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 3601 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 3661 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 3721 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 3781 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 3841 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 3901 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 3961 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 4021 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 4081 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 4141 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 4201 cggcatggac gagctgtaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg 4261 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa 4321 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg 4381 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa 4441 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat 4501 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt 4561 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa 4621 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg 4681 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt 4741 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctaggccacc atggctgaac 4801 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 4861 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 4921 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 4981 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 5041 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 5101 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 5161 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 5221 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 5281 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 5341 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 5401 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 5461 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 5521 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 5581 tctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 5641 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 5701 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 5761 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 5821 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 5881 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 5941 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 6001 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 6061 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 6121 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 6181 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 6241 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct 6301 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 6361 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 6421 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 6481 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 6541 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat 6601 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 6661 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca 6721 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 6781 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact 6841 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 6901 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 6961 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 7021 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 7081 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 7141 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 7201 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 7261 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 7321 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 7381 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 7441 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 7501 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 7561 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 7621 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 7681 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 7741 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 7801 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 7861 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 7921 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 7981 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 8041 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 8101 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 8161 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 8221 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 8281 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 8341 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 8401 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 8461 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 8521 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 8581 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 8641 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 8701 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 8761 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 8821 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 8881 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 8941 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 9001 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 9061 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa 9121 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 9181 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact 9241 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca 9301 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt 9361 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg 9421 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga 9481 cggccagtgc caagctg //