LOCUS V029544 9556 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029544 VERSION V029544 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..4283 /label="NR5A1(NM_004959)" /note="NR5A1(NM_004959)" /gene="NR5A1" promoter 4344..4843 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4864..5463 /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 5477..6065 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6272..6905 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7034..7050) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7058..7074 /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(7082..7112) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7127..7148 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7436..8024) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8195..9055) /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(9056..9160) /gene="bla" /label="AmpR promoter" polyA_signal 9208..9342 /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 caagttcgaa atggactatt cgtacgacga ggacctggac gagctgtgcc 2941 ccgtgtgcgg ggacaaggtg tccggctacc actacggact gctcacgtgt gagagctgca 3001 agggcttctt caagcgcacg gtgcagaaca acaagcacta cacgtgcacc gagagccaga 3061 gctgcaagat cgacaagacg cagcgcaagc gctgtccctt ctgccgcttc cagaaatgcc 3121 tgacggtggg gatgcgcctg gaagccgtgc gcgctgaccg tatgaggggt ggccggaaca 3181 agtttgggcc gatgtacaag cgggaccggg ccctgaaaca gcagaagaag gcacagattc 3241 gggccaatgg cttcaagctg gagacagggc ccccgatggg ggtgcccccg ccgccccctc 3301 ccgcaccgga ctacgtgctg cctcccagcc tgcatgggcc tgagcccaag ggcctggccg 3361 ccggtccacc tgctgggcca ctgggcgact ttggggcccc agcactgccc atggccgtgc 3421 ccggtgccca cgggccactg gctggctacc tctaccctgc ctttcctggc cgtgccatca 3481 agtctgagta cccggagcct tatgccagcc ccccacagcc tgggctgccg tacggctacc 3541 cagagccctt ctctggaggg cccaacgtgc ctgagctcat cctgcagctg ctgcagctgg 3601 agccggatga ggaccaggtg cgggcccgca tcttgggctg cctgcaggag cccaccaaaa 3661 gccgccccga ccagccggcg gccttcggcc tcctgtgcag aatggccgac cagaccttca 3721 tctccatcgt ggactgggca cgcaggtgca tggtcttcaa ggagctggag gtggccgacc 3781 agatgacgct gctgcagaac tgctggagcg agctgctggt gttcgaccac atctaccgcc 3841 aggtccagca cggcaaggag ggcagcatcc tgctggtcac cgggcaggag gtggagctga 3901 ccacagtggc cacccaggcg ggctcgctgc tgcacagcct ggtgttgcgg gcgcaggagc 3961 tggtgctgca gctgcttgcg ctgcagctgg accggcagga gtttgtctgc ctcaagttca 4021 tcatcctctt cagcctggat ttgaagttcc tgaataacca catcctggtg aaagacgctc 4081 aggagaaggc caacgccgcc ctgcttgact acaccctgtg ccactacccg cactgcgggg 4141 acaaattcca gcagctgctg ctgtgcctgg tggaggtgcg ggccctgagc atgcaggcca 4201 aggagtacct gtaccacaag cacctgggca acgagatgcc ccgcaacaac ctgctcatcg 4261 aaatgctgca agccaagcag acttgaattc tgcagtcgac ggtaccgcgg gcccgggatc 4321 ccgcgactct agataattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc 4381 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac 4441 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc 4501 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag 4561 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga 4621 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt 4681 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg 4741 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac 4801 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt 4861 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 4921 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 4981 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 5041 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 5101 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 5161 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5221 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5281 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5341 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5401 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 5461 tgaccgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 5521 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 5581 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 5641 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 5701 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 5761 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 5821 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc 5881 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta 5941 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 6001 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 6061 cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc 6121 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg 6181 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat 6241 cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga 6301 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac 6361 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta 6421 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac 6481 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac 6541 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc 6601 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg 6661 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg 6721 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 6781 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 6841 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 6901 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 6961 atcagagagt gagaggcctt gacattgcta gcgttttacc gtcgacctct agctagagct 7021 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 7081 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 7141 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 7201 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 7261 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 7321 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 7381 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 7441 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 7501 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 7561 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 7621 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 7681 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 7741 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 7801 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 7861 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 7921 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 7981 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 8041 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 8101 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 8161 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 8221 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 8281 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 8341 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 8401 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 8461 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 8521 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 8581 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 8641 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 8701 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 8761 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 8821 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 8881 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 8941 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 9001 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 9061 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 9121 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 9181 cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa 9241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 9301 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct 9361 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg 9421 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt 9481 tgttaaatat gtactacaaa cttagtagtt tttaaagaaa ttgtatttgt taaatatgta 9541 ctacaaactt agtagt //