LOCUS V020751 9946 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020751 VERSION V020751 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 2843..3559 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 3629..4714 /label="KLF1(NM_006563)" /note="KLF1(NM_006563)" /gene="KLF1" promoter 4734..5233 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5254..5853 /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 5867..6455 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6662..7295 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7424..7440) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7448..7464 /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(7472..7502) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7517..7538 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7826..8414) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8585..9445) /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(9446..9550) /gene="bla" /label="AmpR promoter" polyA_signal 9598..9732 /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 ctcaagcttc gaattcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 3241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 3541 catggacgag ctgtacaagt ccggactcag atctcgaggc ggcggaggga gtggaggagg 3601 gggaagcgga ggcggaggga gtggatccat ggccacagcc gagaccgcct tgccctccat 3661 cagcacactg accgccctgg gccccttccc ggacacacag gatgacttcc tcaagtggtg 3721 gcgctccgaa gaggcgcagg acatgggccc gggtcctcct gaccccacgg agccgcccct 3781 ccacgtgaag tctgaggacc agcccgggga ggaagaggac gatgagaggg gcgcggacgc 3841 cacctgggac ctggatctcc tcctcaccaa cttctcgggc ccggagcccg gtggcgcgcc 3901 ccagacctgc gctctggcgc ccagcgaggc ctccggggcg caatatccgc cgccgcccga 3961 gactctgggc gcatatgctg gcggcccggg gctggtggct gggcttttgg gttcggagga 4021 tcactcgggt tgggtgcgcc ctgccctgcg agcccgggct cccgacgcct tcgtgggccc 4081 agccctggct ccagccccgg cccccgagcc caaggcgctg gcgctgcaac cggtgtaccc 4141 ggggcccggc gccggctcct cgggtggcta cttcccgcgg accgggcttt cagtgcctgc 4201 ggcgtcgggc gccccctacg ggctactgtc cgggtacccc gcgatgtacc cggcgcctca 4261 gtaccaaggg cacttccagc tcttccgcgg gctccaggga cccgcgcccg gtcccgccac 4321 gtccccctcc ttcctgagtt gtttgggacc cgggacggtg ggcactggac tcggggggac 4381 tgcagaggat ccaggtgtga tagccgagac cgcgccatcc aagcgaggcc gacgttcgtg 4441 ggcgcgcaag aggcaggcag cgcacacgtg cgcgcacccg ggttgcggca agagctacac 4501 caagagctcc cacctgaagg cgcatctgcg cacgcacaca ggggagaagc catacgcctg 4561 cacgtgggaa ggctgcggct ggagattcgc gcgctcggac gagctgaccc gccactaccg 4621 gaaacacacg gggcagcgcc ccttccgctg ccagctctgc ccacgtgctt tttcgcgctc 4681 tgaccacctg gccttgcaca tgaagcgcca cctttaatct agataattct accgggtagg 4741 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 4801 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 4861 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 4921 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 4981 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 5041 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 5101 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg 5161 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc 5221 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc 5281 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 5341 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 5401 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 5461 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 5521 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 5581 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 5641 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 5701 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 5761 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc 5821 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga 5881 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 5941 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 6001 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 6061 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 6121 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 6181 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 6241 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 6301 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 6361 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 6421 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac 6481 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 6541 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 6601 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga 6661 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 6721 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 6781 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 6841 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 6901 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 6961 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 7021 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 7081 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 7141 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 7201 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 7261 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 7321 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta 7381 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 7441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 7501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 7561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 7621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 7681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 7741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 8041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 8101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 8161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 8221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 8281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 8341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 8401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 8461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 8521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 8581 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 8641 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 8701 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 8761 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 8821 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 8881 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 8941 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 9001 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 9061 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 9121 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 9181 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 9241 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 9301 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 9361 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 9421 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 9481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 9541 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 9601 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 9661 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 9721 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 9781 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 9841 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 9901 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //