LOCUS V041224 6747 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041224 VERSION V041224 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 61..364 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 365..568 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 641..2599 /label="PIAS1(NM_001320687)" /note="PIAS1(NM_001320687)" /gene="PIAS1" CDS 2600..2617 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 2618..2671 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 2693..3412 /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" polyA_signal 3535..3656 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(3663..4118) /direction=LEFT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4145..4249 /gene="bla" /label="AmpR promoter" promoter 4251..4608 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4643..5437 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5669..5716 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 6045..6633 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagct caagcttcga attcgccacc atgtttacac tccaagattc 661 gtatgttaag caaatggtta tgagccttag agtttctgaa ctccaagtac tgttgggcta 721 cgccgggaga aacaagcacg gacgcaaaca cgaacttctc acaaaagccc tgcatttgct 781 aaaggctggc tgtagtcctg ctgtgcaaat gaaaattaag gaactctata ggcggcggtt 841 cccacagaaa atcatgacgc ctgcagactt gtccatcccc aacgtacatt caagtcctat 901 gccagcaact ttgtctccat ctaccattcc acaactcact tacgatggtc accctgcatc 961 atcgccatta ctccctgttt ctcttctggg acctaaacat gaactggaac tcccacatct 1021 tacatcagct cttcacccag tccatccgga tataaaactt caaaaattac cattttatga 1081 tttactggat gaactgataa aacccaccag tctagcatca gacaacagtc agcgctttcg 1141 agaaacctgt tttgcatttg ccttgacacc acaacaagtg cagcaaatca gtagttccat 1201 ggatatttct gggaccaaat gtgacttcac agtacaggtc cagttaaggt tttgtttatc 1261 agaaaccagt tgtccacaag aagatcactt cccacccaat ctttgtgtga aagtgaatac 1321 aaaaccttgc agccttccag gttaccttcc acctacaaaa aatggcgtgg aaccaaagcg 1381 acccagccga ccaattaata tcacctcact tgtccgactg tccacaacag taccaaacac 1441 gattgttgtt tcttggactg cagaaattgg aagaaactat tccatggcag tatatcttgt 1501 aaaacagttg tcctcaacag ttcttcttca gaggttacga gcaaagggaa taaggaatcc 1561 ggatcattct agagctttaa ttaaagagaa gttgactgcg gatccagaca gtgaaatagc 1621 tacaaccagc ctaagggttt ctctactatg tccacttggt aaaatgcggc tgacaattcc 1681 gtgtcgggcc cttacatgtt ctcatctaca atgttttgac gcaactcttt acattcagat 1741 gaatgagaaa aaaccaacct gggtttgtcc tgtctgtgat aagaaggctc catatgaaca 1801 ccttattatt gatggcttgt ttatggaaat cctaaagtac tgtacagact gtgatgaaat 1861 acaatttaag gaggatggca cttgggcacc gatgagatca aaaaaggaag tacaggaagt 1921 ttctgcctct tacaatggag tcgatggatg cttgagctcc acattggagc atcaggtagc 1981 gtctcaccac cagtcctcaa ataaaaacaa gaaagtagaa gtgattgacc taaccataga 2041 cagttcatct gatgaagagg aagaagagcc atctgccaag aggacctgtc cttccctatc 2101 tcccacatca ccactaaata ataaaggcat tttaagtctt ccacatcaag catctccagt 2161 atcccgcacc ccaagccttc ctgctgtaga cacaagctac attaatacct ccctcatcca 2221 agactatagg catcctttcc acatgacacc catgccttac gacttacaag gattagattt 2281 ctttcctttc ttatcaggag acaatcagca ttacaacacc tccttgcttg ccgctgcagc 2341 agcagcagtt tcagatgatc aagacctcct acactcgtct cggtttttcc cgtatacctc 2401 ctcacagatg tttcttgatc agttaagtgc aggaggcagt acttctctgc caaccaccaa 2461 tggaagcagt agtggcagta acagcagcct ggtttcttcc aacagcctaa gggaaagcca 2521 tagccacacc gtcacaaaca ggagcagcac ggacacggca tccatctttg gcatcatacc 2581 agacattatt tcattggacc atcaccatca ccatcacgag ggcagaggaa gtcttctaac 2641 atgcggtgac gtggaggaga atcccggccc tgcggatcca ccggtcgcca ccatggtgag 2701 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 2761 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 2821 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 2881 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 2941 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 3001 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 3061 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 3121 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 3181 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 3241 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 3301 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 3361 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg 3421 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 3481 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 3541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 3601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc 3661 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 3721 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 3781 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 3841 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 3901 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 3961 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 4021 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 4081 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa 4141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 4201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga 4261 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 4321 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 4561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga 4621 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 4681 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 4741 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 4801 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 4861 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 4921 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 4981 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 5041 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 5101 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 5161 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 5221 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 5281 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 5341 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 5401 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 5461 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 5521 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 5581 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag 5641 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt 5701 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc 5761 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc 5821 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata 5881 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 5941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 6001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 6061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 6121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 6181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 6241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 6301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 6361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 6421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 6481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 6541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 6601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 6661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 6721 gtggataacc gtattaccgc catgcat //