LOCUS V018283 10086 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018283 VERSION V018283 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 3545..4879 /label="NEK2(NM_002497)" /note="NEK2(NM_002497)" /gene="NEK2" promoter 4921..5420 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5441..6040 /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 6054..6642 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6849..7482 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7611..7627) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7635..7651 /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(7659..7689) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7704..7725 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8013..8601) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8772..9632) /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(9633..9737) /gene="bla" /label="AmpR promoter" polyA_signal 9785..9919 /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 agtcatgcct tcccgggctg aggactatga agtgttgtac accattggca caggctccta 3601 cggccgctgc cagaagatcc ggaggaagag tgatggcaag atattagttt ggaaagaact 3661 tgactatggc tccatgacag aagctgagaa acagatgctt gtttctgaag tgaatttgct 3721 tcgtgaactg aaacatccaa acatcgttcg ttactatgat cggattattg accggaccaa 3781 tacaacactg tacattgtaa tggaatattg tgaaggaggg gatctggcta gtgtaattac 3841 aaagggaacc aaggaaaggc aatacttaga tgaagagttt gttcttcgag tgatgactca 3901 gttgactctg gccctgaagg aatgccacag acgaagtgat ggtggtcata ccgtattgca 3961 tcgggatctg aaaccagcca atgttttcct ggatggcaag caaaacgtca agcttggaga 4021 ctttgggcta gctagaatat taaaccatga cacgagtttt gcaaaaacat ttgttggcac 4081 accttattac atgtctcctg aacaaatgaa tcgcatgtcc tacaatgaga aatcagatat 4141 ctggtcattg ggctgcttgc tgtatgagtt atgtgcatta atgcctccat ttacagcttt 4201 tagccagaaa gaactcgctg ggaaaatcag agaaggcaaa ttcaggcgaa ttccataccg 4261 ttactctgat gaattgaatg aaattattac gaggatgtta aacttaaagg attaccatcg 4321 accttctgtt gaagaaattc ttgagaaccc tttaatagca gatttggttg cagacgagca 4381 aagaagaaat cttgagagaa gagggcgaca attaggagag ccagaaaaat cgcaggattc 4441 cagccctgta ttgagtgagc tgaaactgaa ggaaattcag ttacaggagc gagagcgagc 4501 tctcaaagca agagaagaaa gattggagca gaaagaacag gagctttgtg ttcgtgagag 4561 actagcagag gacaaactgg ctagagcaga aaatctgttg aagaactaca gcttgctaaa 4621 ggaacggaag ttcctgtctc tggcaagtaa tccagaactt cttaatcttc catcctcagt 4681 aattaagaag aaagttcatt tcagtgggga aagtaaagag aacatcatga ggagtgagaa 4741 ttctgagagt cagctcacat ctaagtccaa gtgcaaggac ctgaagaaaa ggcttcacgc 4801 tgcccagctg cgggctcaag ccctgtcaga tattgagaaa aattaccaac tgaaaagcag 4861 acagatcctg ggcatgcgct agggatccac cggatctaga taactgatca taattctacc 4921 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg 4981 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc 5041 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca 5101 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac 5161 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt 5221 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg 5281 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct 5341 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt 5401 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac 5461 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 5521 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 5581 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 5641 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 5701 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 5761 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 5821 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 5881 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 5941 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 6001 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac 6061 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 6121 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 6181 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 6241 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 6301 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 6361 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 6421 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg 6481 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 6541 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 6601 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag 6661 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga 6721 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt 6781 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 6841 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat 6901 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag 6961 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga 7021 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga 7081 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt 7141 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg 7201 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc 7261 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc 7321 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 7381 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 7441 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc 7501 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac 7561 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 7621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 7681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 7741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 7801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 7861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 7921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 7981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 8041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 8101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 8161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 8221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 8281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 8341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 8401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 8461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 8521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 8581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 8641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 8701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 8761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 8821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 8881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 8941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 9001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 9061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 9121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 9181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 9241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 9301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 9361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 9421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 9481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 9541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 9601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 9661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 9721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga 9781 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 9841 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 9901 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc 9961 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga 10021 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt 10081 agtagt //