LOCUS Exported 5567 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Tag5Amyc-GSK3b WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5567) AUTHORS Zhou BP, Deng J, Xia W, Xu J, Li YM, Gunduz M, Hung MC TITLE Dual regulation of Snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition. JOURNAL Nat Cell Biol. 2004 Oct . 6(10):931-40. PUBMED 15448698 REFERENCE 2 (bases 1 to 5567) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5567 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 124..427 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 428..631 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 582..602 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 677..695 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 1597..1608 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS 2043..2072 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(2125..2144) /label=T7 /note="T7 promoter, forward primer" promoter complement(2126..2144) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2418..2539 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2455..2474) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2509..2528 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2546..3001) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2683..2704) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2895..2914 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3028..3132 /gene="bla" /label=AmpR promoter promoter 3134..3491 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3157..3177) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3342..3477 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3404..3423 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3526..4320 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3580..3599) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4190..4209 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4508..4527) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4552..4599 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4928..5516 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5417..5436 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg 61 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt 121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc 181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg 241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat 301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca 361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat 421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg 481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca 541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg 601 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg 661 attacgccaa gctcgaaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg 721 gtggcggccg ctctagcccg ggcggatccc ccgggctgca ggaattcgat atcgccacca 781 tgtcagggcg gcccagaacc acctcctttg cggagagctg caagccggtg cagcagcctt 841 cagcttttgg cagcatgaaa gttagcagag acaaggacgg cagcaaggtg acaacagtgg 901 tggcaactcc tgggcagggt ccagacaggc cacaagaagt cagctataca gacactaaag 961 tgattggaaa tggatcattt ggtgtggtat atcaagccaa actttgtgat tcaggagaac 1021 tggtcgccat caagaaagta ttgcaggaca agagatttaa gaatcgagag ctccagatca 1081 tgagaaagct agatcactgt aacatagtcc gattgcgtta tttcttctac tccagtggtg 1141 agaagaaaga tgaggtctat cttaatctgg tgctggacta tgttccggaa acagtataca 1201 gagttgccag acactatagt cgagccaaac agacgctccc tgtgatttat gtcaagttgt 1261 atatgtatca gctgttccga agtttagcct atatccattc ctttggaatc tgccatcggg 1321 atattaaacc gcagaacctc ttgttggatc ctgatactgc tgtattaaaa ctctgtgact 1381 ttggaagtgc aaagcagctg gtccgaggag aacccaatgt ttcgtatatc tgttctcggt 1441 actatagggc accagagttg atctttggag ccactgatta tacctctagt atagatgtat 1501 ggtctgctgg ctgtgtgttg gctgagctgt tactaggaca accaatattt ccaggggata 1561 gtggtgtgga tcagttggta gaaataatca aggtcctggg aactccaaca agggagcaaa 1621 tcagagaaat gaacccaaac tacacagaat ttaaattccc tcaaattaag gcacatcctt 1681 ggactaaggt cttccgaccc cgaactccac cggaggcaat tgcactgtgt agccgtctgc 1741 tggagtatac accaactgcc cgactaacac cactggaagc ttgtgcacat tcattttttg 1801 atgaattacg ggacccaaat gtcaaactac caaatgggcg agacacacct gcactcttca 1861 acttcaccac tcaagaactg tcaagtaatc cacctctggc taccatcctt attcctcctc 1921 atgctcggat tcaagcagct gcttcaaccc ccacaaatgc cacagcagcg tcagatgcta 1981 atactggaga ccgtggacag accaataatg ctgcttctgc atcagcttcc aactccaccc 2041 tcgagcagaa actcatctct gaagaggatc tgtagggccc ggtaccttaa ttaattaagg 2101 taccaggtaa gtgtacccaa ttcgccctat agtgagtcgt attacaattc actcgatcgc 2161 ccttcccaac agttgcgcag cctgaatggc gaatggagat ccaattttta agtgtataat 2221 gtgttaaact actgattcta attgtttgtg tattttagat tcacagtccc aaggctcatt 2281 tcaggcccct cagtcctcac agtctgttca tgatcataat cagccatacc acatttgtag 2341 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 2401 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 2461 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 2521 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg 2581 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct 2641 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt 2701 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat 2761 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca 2821 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac 2881 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta 2941 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg 3001 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata 3061 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 3121 aaaaggaaga atcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg 3181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3241 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3301 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 3361 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 3421 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 3481 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 3541 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 3601 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 3661 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg 3721 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3781 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3841 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 3901 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3961 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagaaca tcaggggctc 4021 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc 4081 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 4141 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 4201 cgtgatattg ctgaagaact tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 4261 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 4321 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 4381 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 4441 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga 4501 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 4561 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 4621 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 4681 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 4741 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa 4801 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 4861 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 4921 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 4981 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 5041 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 5101 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 5161 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 5221 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 5281 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 5341 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 5401 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 5461 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 5521 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gcctttt //