LOCUS Exported 5071 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Flag Snail WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5071) 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 5071) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5071 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 16..319 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 320..523 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 474..494 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 569..587 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" regulatory 622..631 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 631..654 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(1521..1540) /label=T7 /note="T7 promoter, forward primer" promoter complement(1522..1540) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 1814..1935 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1851..1870) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1905..1924 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1942..2397) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2079..2100) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2291..2310 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2424..2528 /gene="bla" /label=AmpR promoter promoter 2530..2887 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2553..2573) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2738..2873 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2800..2819 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2922..3716 /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(2976..2995) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3586..3605 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3904..3923) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3948..3995 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4324..4912 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4813..4832 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca 541 gatccgctag cgattacgcc aagctcgaaa ttaaccctca ctaaagggaa caaaagctgg 601 agctccaccg cggtggcggc cgccaccatg gattacaagg atgacgacga taagagcccg 661 ggcggatccc cgcgctcttt cctcgtcagg aagccctccg accccaatcg gaagcctaac 721 tacagcgagc tgcaggactc taatccagag tttaccttcc agcagcccta cgaccaggcc 781 cacctgctgg cagccatccc acctccggag atcctcaacc ccaccgcctc gctgccaatg 841 ctcatctggg actctgtcct ggcgccccaa gcccagccaa ttgcctgggc ctcccttcgg 901 ctccaggaga gtcccagggt ggcagagctg acctccctgt cagatgagga cagtgggaaa 961 ggctcccagc cccccagccc accctcaccg gctccttcgt ccttctcctc tacttcagtc 1021 tcttccttgg aggccgaggc ctatgctgcc ttcccaggct tgggccaagt gcccaagcag 1081 ctggcccagc tctctgaggc caaggatctc caggctcgaa aggccttcaa ctgcaaatac 1141 tgcaacaagg aatacctcag cctgggtgcc ctcaagatgc acatccgaag ccacacgctg 1201 ccctgcgtct gcggaacctg cgggaaggcc ttctctaggc cctggctgct acaaggccat 1261 gtccggaccc acactggcga gaagcccttc tcctgtcccc actgcagccg tgccttcgct 1321 gaccgctcca acctgcgggc ccacctccag acccactcag atgtcaagaa gtaccagtgc 1381 caggcgtgtg ctcggacctt ctcccgaatg tccctgctcc acaagcacca agagtccggc 1441 tgctcaggat gtccccgctg actcgagggg gggcccggta ccttaattaa ttaaggtacc 1501 aggtaagtgt acccaattcg ccctatagtg agtcgtatta caattcactc gatcgccctt 1561 cccaacagtt gcgcagcctg aatggcgaat ggagatccaa tttttaagtg tataatgtgt 1621 taaactactg attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag 1681 gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt 1741 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc 1801 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 1861 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 1921 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 1981 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 2041 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 2101 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 2161 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 2221 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 2281 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 2341 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag 2401 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 2461 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 2521 ggaagaatcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa 2581 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 2641 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 2701 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 2761 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 2821 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 2881 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 2941 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3001 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3061 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc 3121 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 3181 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 3241 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 3301 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 3361 ggatggaagc cggtcttgtc gatcaggatg atctggacga agaacatcag gggctcgcgc 3421 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga 3481 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 3541 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 3601 atattgctga agaacttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 3661 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 3721 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 3781 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 3841 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct 3901 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag 3961 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg 4021 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc 4081 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg 4141 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact 4201 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 4261 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 4321 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 4381 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 4441 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 4501 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 4561 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 4621 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 4681 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 4741 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 4801 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 4861 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 4921 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 4981 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt 5041 aatcaattac ggggtcatta gttcatagcc c //