LOCUS V018564 10746 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018564 VERSION V018564 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" CDS 2807..3523 /codon_start=1 /note="mammalian codon-optimized" /product="Aequorea coerulescens GFP" /transl_table=1 /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF GFVTAAAITHGMDELYK" /label="AcGFP1" CDS 3560..5539 /label="ACOX1(NM_004035)" /note="ACOX1(NM_004035)" /gene="ACOX1" promoter 5581..6080 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6101..6700 /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 6714..7302 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7509..8142 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8271..8287) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8295..8311 /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(8319..8349) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8364..8385 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8673..9261) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9432..10292) /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(10293..10397) /gene="bla" /label="AmpR promoter" polyA_signal 10445..10579 /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 tctaccggtc gccaccatgg tgagcaaggg 2821 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg 2881 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct 2941 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct 3001 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt 3061 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg 3121 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga 3181 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa 3241 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa 3301 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca 3361 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca 3421 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt 3481 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac tcagatctcg 3541 agctcaagct tcgaattcga tgaacccgga cctgcgcagg gagcgggatt ccgccagctt 3601 caacccggag ctgcttacac acatcctgga cggcagcccc gagaaaaccc ggcgccgccg 3661 agagatcgag aacatgatcc tgaacgaccc agacttccag catgaggact tgaacttcct 3721 cactcgcagc cagcgttatg aggtggctgt caggaaaagt gccatcatgg tgaagaagat 3781 gagggagttt ggcatcgctg accctgatga aattatgtgg tttaaaaatt ttgtgcaccg 3841 agggcggcct gagcctctgg atcttcactt gggcatgttc ctgcccacct tgcttcacca 3901 ggcaactgcg gagcagcagg agcgcttctt catgcccgcc tggaacttgg agatcattgg 3961 cacttatgcc cagacagaga tgggtcatgg aactcacctt cgaggcttgg aaaccacagc 4021 cacgtatgac cctgaaaccc aggagttcat tctcaacagt cctactgtga cctccattaa 4081 atggtggcct ggtgggcttg gaaagacttc aaatcatgca atagttcttg cccagctcat 4141 cactaagggg aaatgctatg gattacatgc ctttatcgta cctattcgtg aaatcgggac 4201 ccataagcct ttgccaggaa ttaccgttgg tgacatcggc cccaaatttg gttatgatga 4261 gatagacaat ggctacctca aaatggacaa ccatcgtatt cccagagaaa acatgctgat 4321 gaagtatgcc caggtgaagc ctgatggcac atacgtgaaa ccgctgagta acaagctgac 4381 ttacgggacc atggtgtttg tcaggtcctt ccttgtggga gaagctgctc gggctctgtc 4441 taaggcgtgc accattgcca tccgatacag cgctgtgagg caccagtctg aaatcaagcc 4501 aggtgaacca gaaccacaga ttttggattt tcaaacccag cagtataaac tctttccact 4561 cctggccact gcctatgcct tccagtttgt gggcgcatac atgaaggaga cctatcaccg 4621 gattaacgaa ggcattggtc aaggggacct gagtgaactg cctgagcttc atgccctcac 4681 cgctggactg aaggctttca cctcctggac tgcaaacact ggcattgaag catgtcggat 4741 ggcttgtggt gggcatggct attctcattg cagtggtctt ccaaatattt atgtcaattt 4801 caccccaagc tgtacctttg agggagaaaa cactgtcatg atgctccaga cggctaggtt 4861 cctgatgaaa agttatgatc aggtgcactc aggaaagttg gtgtgtggca tggtgtccta 4921 tttgaacgac ctgcccagtc agcgcatcca gccacagcag gtagcagtct ggccaaccat 4981 ggtggatatc aacagccccg aaagcctaac cgaagcatat aaactccgtg cagccagatt 5041 agtagaaatt gctgcaaaaa accttcaaaa agaagtgatt cacagaaaaa gcaaggaggt 5101 agcttggaac ctaacttctg ttgaccttgt tcgagcaagt gaggcacatt gccactatgt 5161 ggtagttaag ctcttttcag aaaaactcct caaaattcaa gataaagcca ttcaagctgt 5221 cttaaggagt ttatgtctgc tgtattctct gtatggaatc agtcagaacg cgggggattt 5281 ccttcagggg agcatcatga cagagcctca gattacacaa gtaaaccagc gtgtaaagga 5341 gttactcact ctgattcgct cagatgctgt tgctttggtt gatgcatttg attttcagga 5401 tgtgacactt ggctctgtgc ttggccgcta tgatgggaat gtgtatgaaa acttgtttga 5461 gtgggctaag aactccccac tgaacaaagc agaggtccac gaatcttaca agcacctgaa 5521 gtcactgcag tccaagctct gaggatccac cggatctaga taactgatca taattctacc 5581 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg 5641 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc 5701 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca 5761 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac 5821 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt 5881 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg 5941 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct 6001 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt 6061 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac 6121 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 6181 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 6241 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 6301 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 6361 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 6421 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 6481 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 6541 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 6601 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 6661 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac 6721 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 6781 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 6841 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 6901 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 6961 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 7021 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 7081 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg 7141 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 7201 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 7261 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag 7321 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga 7381 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt 7441 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 7501 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat 7561 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag 7621 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga 7681 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga 7741 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt 7801 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg 7861 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc 7921 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc 7981 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 8041 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 8101 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc 8161 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac 8221 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 8281 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 8341 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 8401 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 8461 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 8521 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 8581 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 8641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 8701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 8761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 8821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 8881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 8941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 9001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 9061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 9121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 9181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 9241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 9301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 9361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 9421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 9481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 9541 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 9601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 9661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 9721 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 9781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 9841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 9901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 9961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 10021 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 10081 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 10141 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 10201 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 10261 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 10321 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 10381 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga 10441 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 10501 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 10561 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc 10621 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga 10681 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt 10741 agtagt //