LOCUS V036244 6930 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036244 VERSION V036244 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1055..1771 /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 1808..3106 /label="GSK3B(NM_002093)" /note="GSK3B(NM_002093)" /gene="GSK3B" CDS 3113..3178 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" primer_bind complement(3187..3203) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 3214..3836 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3985..4573) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 4902..4949 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(5181..5975) /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" promoter complement(6010..6367) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(6369..6473) /gene="bla" /label="AmpR promoter" rep_origin 6500..6928 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgcggc gcgccagatc tcaagcttgc caccatggtg agcaagggcg aggagctgtt 1081 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 1141 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 1201 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 1261 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 1321 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 1381 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 1441 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 1501 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 1561 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 1621 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 1681 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 1741 gatcactctc ggcatggacg agctgtacaa gggcggtgga ggcagtggtg gaggcggtag 1801 ctctagaatg tcagggcggc ccagaaccac ctcctttgcg gagagctgca agccggtgca 1861 gcagccttca gcttttggca gcatgaaagt tagcagagac aaggacggca gcaaggtgac 1921 aacagtggtg gcaactcctg ggcagggtcc agacaggcca caagaagtca gctatacaga 1981 cactaaagtg attggaaatg gatcatttgg tgtggtatat caagccaaac tttgtgattc 2041 aggagaactg gtcgccatca agaaagtatt gcaggacaag agatttaaga atcgagagct 2101 ccagatcatg agaaagctag atcactgtaa catagtccga ttgcgttatt tcttctactc 2161 cagtggtgag aagaaagatg aggtctatct taatctggtg ctggactatg ttccggaaac 2221 agtatacaga gttgccagac actatagtcg agccaaacag acgctccctg tgatttatgt 2281 caagttgtat atgtatcagc tgttccgaag tttagcctat atccattcct ttggaatctg 2341 ccatcgggat attaaaccgc agaacctctt gttggatcct gatactgctg tattaaaact 2401 ctgtgacttt ggaagtgcaa agcagctggt ccgaggagaa cccaatgttt cgtatatctg 2461 ttctcggtac tatagggcac cagagttgat ctttggagcc actgattata cctctagtat 2521 agatgtatgg tctgctggct gtgtgttggc tgagctgtta ctaggacaac caatatttcc 2581 aggggatagt ggtgtggatc agttggtaga aataatcaag gtcctgggaa ctccaacaag 2641 ggagcaaatc agagaaatga acccaaacta cacagaattt aaattccctc aaattaaggc 2701 acatccttgg actaaggatt cgtcaggaac aggacatttc acctcaggag tgcgggtctt 2761 ccgaccccga actccaccgg aggcaattgc actgtgtagc cgtctgctgg agtatacacc 2821 aactgcccga ctaacaccac tggaagcttg tgcacattca ttttttgatg aattacggga 2881 cccaaatgtc aaactaccaa atgggcgaga cacacctgca ctcttcaact tcaccactca 2941 agaactgtca agtaatccac ctctggctac catccttatt cctcctcatg ctcggattca 3001 agcagctgct tcaaccccca caaatgccac agcagcgtca gatgctaata ctggagaccg 3061 tggacagacc aataatgctg cttctgcatc agcttccaac tccaccggat ccgactacaa 3121 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 3181 gccgcggtca tagctgtttc ctgaacagat cccgggtggc atccctgtga cccctcccca 3241 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa 3301 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg 3361 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg 3421 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt 3481 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg 3541 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc 3601 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag 3661 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag 3721 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc 3781 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt 3841 acgcggccag cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 3901 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 3961 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 4021 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 4081 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 4141 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 4201 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 4261 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 4321 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 4381 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 4441 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 4501 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 4561 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 4621 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 4681 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat 4741 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg 4801 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt 4861 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc 4921 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc 4981 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc 5041 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata 5101 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat 5161 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc 5221 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca 5281 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcgatc cgccacaccc 5341 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag 5401 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg 5461 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca 5521 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat 5581 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact 5641 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc 5701 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc 5761 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg 5821 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca 5881 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc 5941 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct 6001 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag 6061 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg 6121 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac 6181 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg 6241 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc 6301 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc 6361 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 6421 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 6481 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 6541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 6601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 6661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 6721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 6781 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 6841 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 6901 ctgatttaac aaaaatttaa cgcgaatttt //