LOCUS Exported 7641 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3 deltaN47 Beta-catenin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7641) AUTHORS Kolligs FT, Hu G, Dang CV, Fearon ER TITLE Neoplastic transformation of RK3E by mutant beta-catenin requires deregulation of Tcf/Lef transcription but not activation of c-myc expression. JOURNAL Mol Cell Biol. 1999 Aug . 19(8):5696-706. PUBMED 10409758 REFERENCE 2 (bases 1 to 7641) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7641 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(63..80) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(314..333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(993..1853) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1616..1635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1854..1958) /gene="bla" /label=AmpR promoter primer_bind complement(2033..2052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2224..2603 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2604..2807 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2758..2778 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2852..2871 /label=T7 /note="T7 promoter, forward primer" promoter 2852..2870 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 5122..5145 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(5183..5201) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5184..5201) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5221..5238) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5227..5451 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5497..5925 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5584..5603) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5794..5815 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5934..5954) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5939..6268 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6119..6254 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6181..6200 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6335..7129 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6389..6408) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6999..7018 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 7303..7424 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7340..7359) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7394..7413 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7473..7489) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7473..7489) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7486..7508) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7497..7513 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(7521..7551) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7566..7587 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 2881 cttggtaccg agctcggatc cccagcgtgg acaatggctg gtaaaggcaa tcctgaggaa 2941 gaggatgtgg atacctccca agtcctgtat gagtgggaac agggattttc tcagtccttc 3001 actcaagaac aagtagctga tattgatgga cagtatgcaa tgactcgagc tcagagggta 3061 cgagctgcta tgttccctga gacattagat gagggcatgc agatcccatc tacacagttt 3121 gatgctgctc atcccactaa tgtccagcgt ttggctgaac catcacagat gctgaaacat 3181 gcagttgtaa acttgattaa ctatcaagat gatgcagaac ttgccacacg tgcaatccct 3241 gaactgacaa aactgctaaa tgacgaggac caggtggtgg ttaataaggc tgcagttatg 3301 gtccatcagc tttctaaaaa ggaagcttcc agacacgcta tcatgcgttc tcctcagatg 3361 gtgtctgcta ttgtacgtac catgcagaat acaaatgatg tagaaacagc tcgttgtacc 3421 gctgggacct tgcataacct ttcccatcat cgtgagggct tactggccat ctttaagtct 3481 ggaggcattc ctgccctggt gaaaatgctt ggttcaccag tggattctgt gttgttttat 3541 gccattacaa ctctccacaa ccttttatta catcaagaag gagctaaaat ggcagtgcgt 3601 ttagctggtg ggctgcagaa aatggttgcc ttgctcaaca aaacaaatgt taaattcttg 3661 gctattacga cagactgcct tcaaatttta gcttatggca accaagaaag caagctcatc 3721 atactggcta gtggtggacc ccaagcttta gtaaatataa tgaggaccta tacttacgaa 3781 aaactactgt ggaccacaag cagagtgctg aaggtgctat ctgtctgctc tagtaataag 3841 ccggctattg tagaagctgg tggaatgcaa gctttaggac ttcacctgac agatccaagt 3901 caacgtcttg ttcagaactg tctttggact ctcaggaatc tttcagatgc tgcaactaaa 3961 caggaaggga tggaaggtct ccttgggact cttgttcagc ttctgggttc agatgatata 4021 aatgtggtca cctgtgcagc tggaattctt tctaacctca cttgcaataa ttataagaac 4081 aagatgatgg tctgccaagt gggtggtata gaggctcttg tgcgtactgt ccttcgggct 4141 ggtgacaggg aagacatcac tgagcctgcc atctgtgctc ttcgtcatct gaccagccga 4201 caccaagaag cagagatggc ccagaatgca gttcgccttc actatggact accagttgtg 4261 gttaagctct tacacccacc atcccactgg cctctgataa aggctactgt tggattgatt 4321 cgaaatcttg ccctttgtcc cgcaaatcat gcacctttgc gtgagcaggg tgccattcca 4381 cgactagttc agttgcttgt tcgtgcacat caggataccc agcgccgtac gtccatgggt 4441 gggacacagc agcaatttgt ggagggggtc cgcatggaag aaatagttga aggttgtacc 4501 ggagcccttc acatcctagc tcgggatgtt cacaaccgaa ttgttatcag aggactaaat 4561 accattccat tgtttgtgca gctgctttat tctcccattg aaaacatcca aagagtagct 4621 gcaggggtcc tctgtgaact tgctcaggac aaggaagctg cagaagctat tgaagctgag 4681 ggagccacag ctcctctgac agagttactt cactctagga atgaaggtgt ggcgacatat 4741 gcagctgctg ttttgttccg aatgtctgag gacaagccac aagattacaa gaaacggctt 4801 tcagttgagc tgaccagctc tctcttcaga acagagccaa tggcttggaa tgagactgct 4861 gatcttggac ttgatattgg tgcccaggga gaaccccttg gatatcgcca ggatgatcct 4921 agctatcgtt cttttcactc tggtggatat ggccaggatg ccttgggtat ggaccccatg 4981 atggaacatg agatgggtgg ccaccaccct ggtgctgact atccagttga tgggctgcca 5041 gatctggggc atgcccagga cctcatggat gggctgcctc caggtgacag caatcagctg 5101 gcctggtttg atactgacct ggactacaag gacgacgatg acaagtaagc ggccgctcga 5161 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag 5221 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 5281 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 5341 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 5401 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 5461 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 5521 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 5581 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 5641 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 5701 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 5761 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 5821 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 5881 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 5941 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 6001 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 6061 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 6121 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 6181 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 6241 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 6301 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 6361 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 6421 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 6481 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 6541 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 6601 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 6661 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 6721 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 6781 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 6841 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 6901 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 6961 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 7021 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 7081 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 7141 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 7201 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 7261 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 7321 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 7381 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 7441 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 7501 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 7561 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 7621 ggaaacctgt cgtgccagct g //