LOCUS V031798 10006 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031798 VERSION V031798 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 3572..4768 /label="B4galt1(NM_022305)" /note="B4galt1(NM_022305)" /gene="B4galt1" promoter 4841..5340 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5361..5960 /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 5974..6562 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6769..7402 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7531..7547) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7555..7571 /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(7579..7609) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7624..7645 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7933..8521) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8692..9552) /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(9553..9657) /gene="bla" /label="AmpR promoter" polyA_signal 9705..9839 /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 tcagaggagg 3541 tggaggtagt ggaggaggcg gaagcctcga gatgaggttt cgtgagcagt tcctgggcgg 3601 cagcgccgcg atgccgggcg cgaccctgca gcgggcctgc cgcctgctcg tggccgtctg 3661 cgcgctgcac ctcggcgtca ccctcgtcta ttacctctct ggccgcgatc tgagccgcct 3721 gccccagttg gtcggagtct cctctacact gcagggcggc acgaacggcg ccgcagccag 3781 caagcagccc ccaggagagc agcggccgcg gggtgcgcgg ccgccgcctc ctttaggcgt 3841 ctccccgaag cctcgcccgg gtctcgactc cagccctggt gcagcttctg gccccggctt 3901 gaagagcaac ttgtcttcgt tgccagtgcc caccaccact ggactgttgt cgctgccagc 3961 ttgccctgag gagtccccgc tgctcgttgg ccccatgctg attgacttta atattgctgt 4021 ggatctggag cttttggcaa agaagaaccc agagataaag acgggcggcc gttactcccc 4081 caaggactgt gtctctcctc acaaggtggc catcatcatc ccattccgta accggcagga 4141 gcatctcaaa tactggctgt attatttgca tcccatcctt cagcgccagc aactcgacta 4201 tggcatctac gtcatcaatc aggctggaga caccatgttc aatcgagcta agctgctcaa 4261 tattggcttt caagaggcct tgaaggacta tgattacaac tgctttgtgt tcagtgatgt 4321 ggacctcatt ccgatggacg accgtaatgc ctacaggtgt ttttcgcagc cacggcacat 4381 ttctgttgca atggacaagt tcgggtttag cctgccatat gttcagtatt ttggaggtgt 4441 ctctgctctc agtaaacaac agtttcttgc catcaatgga ttccctaata attattgggg 4501 ttggggagga gaagatgacg acatttttaa cagattagtt cataaaggca tgtctatatc 4561 acgtccaaat gctgtagtag ggaggtgtcg aatgatccgg cattcaagag acaagaaaaa 4621 tgagcccaat cctcagaggt ttgaccggat cgcacataca aaggaaacga tgcgcttcga 4681 tggtttgaac tcacttacct acaaggtgtt ggatgtacag agatacccgt tatataccca 4741 aatcacagtg gacatcggga caccgagata ggaattctgc agtcgacggt accgcgggcc 4801 cgggatccac cggatctaga taactgatca taattctacc gggtagggga ggcgcttttc 4861 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt 4921 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt 4981 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg 5041 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag 5101 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag 5161 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc 5221 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc 5281 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg 5341 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg 5401 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc 5461 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc 5521 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg 5581 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc 5641 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg 5701 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc 5761 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg 5821 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc 5881 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga 5941 cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt 6001 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct 6061 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat 6121 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg 6181 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag 6241 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc 6301 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg 6361 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc 6421 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc 6481 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc 6541 tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg 6601 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac 6661 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt 6721 acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa 6781 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact 6841 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat 6901 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga 6961 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt 7021 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg 7081 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc 7141 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag 7201 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc 7261 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa 7321 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag 7381 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa 7441 cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc 7501 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 7561 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 7621 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 7681 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 7741 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 7801 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 7861 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 7921 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7981 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 8041 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 8101 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 8161 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 8221 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 8281 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 8341 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 8401 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 8461 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 8521 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 8581 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 8641 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 8701 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 8761 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 8821 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 8881 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 8941 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 9001 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 9061 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 9121 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 9181 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 9241 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 9301 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 9361 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 9421 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 9481 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 9541 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 9601 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 9661 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag 9721 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 9781 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca 9841 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca 9901 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat 9961 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //