LOCUS V031808 10207 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031808 VERSION V031808 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..4969 /label="Sox10(NM_011437)" /note="Sox10(NM_011437)" /gene="Sox10" promoter 5042..5541 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5562..6161 /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 6175..6763 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6970..7603 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7732..7748) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7756..7772 /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(7780..7810) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7825..7846 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8134..8722) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8893..9753) /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(9754..9858) /gene="bla" /label="AmpR promoter" polyA_signal 9906..10040 /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 gatggccgag gaacaagacc tatcagaggt 3601 ggagctgagc cctgtgggct cggaggaacc ccgctgcctg tccccaggca gcgcgccgtc 3661 gctgggaccc gacggcggcg gcggtggctc gggcttgcga gccagcccgg ggcccggtga 3721 actgggcaag gtcaagaagg aacagcagga cggcgaggcg gacgatgaca agttccccgt 3781 gtgcatccgc gaggcggtca gccaggtgct cagcggctac gactggacgc tggtgcccat 3841 gcccgtgcgc gtcaacggtg ccagcaagag caagccgcac gtcaagaggc ccatgaacgc 3901 cttcatggtg tgggcacagg cggcacgcag aaagctagcc gaccagtacc ctcacctcca 3961 caatgctgag ctcagcaaga cactaggcaa gctctggagg ttgctgaacg aaagtgacaa 4021 gcgccccttc attgaggagg ctgagaggct ccggatgcag cacaaaaagg accatccgga 4081 ctacaagtac caacctcggc ggcggaagaa cgggaaggca gcccaggggg aggcagaatg 4141 cccaggcggg gaagccgagc aaggaggggc tgctgctatt caggctcact acaagagtgc 4201 ccacctggac caccggcacc cagaagaagg ctcccccatg tcagatggga acccagagca 4261 cccctcaggc cagagccatg gccccccaac ccctccaacc accccaaaga cagagctgca 4321 gtccggcaag gcagacccca aaagggatgg gcgctccttg ggggagggcg ggaagcccca 4381 catcgacttc ggcaacgtgg acatcgggga gatcagccac gaggtaatgt ccaacatgga 4441 aacctttgat gtgactgagc tggaccaata cctgccaccc aacgggcacc caggccatgt 4501 gggtagctac tcggcagctg gctacgggct gggcagtgcc ctggctgtgg ccagtggaca 4561 ctctgcctgg atctccaagc caccaggtgt ggctctgccc acggtctcgc cccctggtgt 4621 ggatgccaaa gcccaggtga agacagagac cacaggcccc cagggacccc cacactacac 4681 cgaccagccg tccacttccc agatcgccta cacttccctc agtctgcccc actacggctc 4741 cgccttcccc tccatctcac gaccccagtt tgactattct gaccatcagc cctcaggacc 4801 ctattatggc catgcaggcc aggcctctgg cctctattca gccttttctt acatggggcc 4861 ctcccagcgg cccctctaca ctgccatctc tgaccccagc ccctcagggc cccaatccca 4921 cagtcccaca cactgggagc agccagtata tacgactcta tcccgacctt aggaattctg 4981 cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc ataattctac 5041 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg 5101 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg 5161 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc 5221 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca 5281 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt 5341 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg 5401 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc 5461 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct 5521 tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca 5581 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 5641 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 5701 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 5761 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 5821 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 5881 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 5941 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 6001 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 6061 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 6121 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa 6181 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 6241 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 6301 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 6361 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 6421 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 6481 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 6541 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt 6601 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 6661 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 6721 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca 6781 gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg 6841 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg 6901 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6961 agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga 7021 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca 7081 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag 7141 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg 7201 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg 7261 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg 7321 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag 7381 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac 7441 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa 7501 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag 7561 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat 7621 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga 7681 cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 7741 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 7801 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 7861 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 7921 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 7981 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 8041 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 8101 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 8161 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 8221 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 8281 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 8341 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 8401 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 8461 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 8521 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 8581 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 8641 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 8701 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 8761 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 8821 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 8881 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 8941 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 9001 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 9061 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 9121 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 9181 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 9241 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 9301 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 9361 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 9421 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 9481 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 9541 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 9601 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 9661 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 9721 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 9781 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 9841 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 9901 agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 9961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 10021 tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc 10081 ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg 10141 attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact 10201 tagtagt //