LOCUS Exported 7681 bp ds-DNA circular SYN 25-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pT3-N90-beta-catenin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7681) AUTHORS Tward AD, Jones KD, Yant S, Cheung ST, Fan ST, Chen X, Kay MA, Wang R, Bishop JM TITLE Distinct pathways of genomic progression to benign and malignant tumors of the liver. JOURNAL Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14771-6. Epub 2007 Sep 4. PUBMED 17785413 REFERENCE 2 (bases 1 to 7681) AUTHORS . TITLE Direct Submission JOURNAL Exported May 25, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7681 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 90..1268 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 321..1259 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1216..1236 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 1285..1304 /label=T7 /note="T7 promoter, forward primer" promoter 1285..1303 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1382..1406 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1417..1446 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" protein_bind complement(3536..3560) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3613..3654 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3692..3709) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3698..3922 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind complement(3992..4025) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind 4081..4101 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" primer_bind complement(4483..4500) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4483..4499) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4492..4514) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(4708..4727) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4827..4849 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4887..4905) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4973..5077 /gene="bla" /label=AmpR promoter CDS 5078..5938 /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 complement(5296..5315) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6109..6697 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6598..6617 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6851..6868 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6985..7006 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7021..7051 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7059..7075 /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)." primer_bind 7064..7086 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7083..7099 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7083..7099 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7525..7544) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind complement(7597..7630) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." ORIGIN 1 aaaaagcttt gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga 61 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca 121 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga 181 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg 241 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg 301 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta 361 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg 421 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc 481 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct 541 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt 601 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc 661 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc 721 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt 781 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct 841 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg 901 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt 961 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg 1021 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt 1081 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg 1141 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg 1201 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg 1261 tgtcgtgagg aattagcttg gtactaatac gactcactat agggagaccc aagctggcta 1321 ggtaagcttg gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc 1381 aacaagtttg tacaaaaaag caggcttcgt accatggagc agaagctgat ctccgaggag 1441 gacctgaagg atcctcgagc tcagagggta cgagctgcta tgttccctga gacattagat 1501 gagggcatgc agatcccatc tacacagttt gatgctgctc atcccactaa tgtccagcgt 1561 ttggctgaac catcacagat gctgaaacat gcagttgtaa acttgattaa ctatcaagat 1621 gatgcagaac ttgccacacg tgcaatccct gaactgacaa aactgctaaa tgacgaggac 1681 caggtggtgg ttaataaggc tgcagttatg gtccatcagc tttctaaaaa ggaagcttcc 1741 agacacgcta tcatgcgttc tcctcagatg gtgtctgcta ttgtacgtac catgcagaat 1801 acaaatgatg tagaaacagc tcgttgtacc gctgggacct tgcataacct ttcccatcat 1861 cgtgagggct tactggccat ctttaagtct ggaggcattc ctgccctggt gaaaatgctt 1921 ggttcaccag tggattctgt gttgttttat gccattacaa ctctccacaa ccttttatta 1981 catcaagaag gagctaaaat ggcagtgcgt ttagctggtg ggctgcagaa aatggttgcc 2041 ttgctcaaca aaacaaatgt taaattcttg gctattacga cagactgcct tcaaatttta 2101 gcttatggca accaagaaag caagctcatc atactggcta gtggtggacc ccaagcttta 2161 gtaaatataa tgaggaccta tacttacgaa aaactactgt ggaccacaag cagagtgctg 2221 aaggtgctat ctgtctgctc tagtaataag ccggctattg tagaagctgg tggaatgcaa 2281 gctttaggac ttcacctgac agatccaagt caacgtcttg ttcagaactg tctttggact 2341 ctcaggaatc tttcagatgc tgcaactaaa caggaaggga tggaaggtct ccttgggact 2401 cttgttcagc ttctgggttc agatgatata aatgtggtca cctgtgcagc tggaattctt 2461 tctaacctca cttgcaataa ttataagaac aagatgatgg tctgccaagt gggtggtata 2521 gaggctcttg tgcgtactgt ccttcgggct ggtgacaggg aagacatcac tgagcctgcc 2581 atctgtgctc ttcgtcatct gaccagccga caccaagaag cagagatggc ccagaatgca 2641 gttcgccttc actatggact accagttgtg gttaagctct tacacccacc atcccactgg 2701 cctctgataa aggctactgt tggattgatt cgaaatcttg ccctttgtcc cgcaaatcat 2761 gcacctttgc gtgagcaggg tgccattcca cgactagttc agttgcttgt tcgtgcacat 2821 caggataccc agcgccgtac gtccatgggt gggacacagc agcaatttgt ggagggggtc 2881 cgcatggaag aaatagttga aggttgtacc ggagcccttc acatcctagc tcgggatgtt 2941 cacaaccgaa ttgttatcag aggactaaat accattccat tgtttgtgca gctgctttat 3001 tctcccattg aaaacatcca aagagtagct gcaggggtcc tctgtgaact tgctcaggac 3061 aaggaagctg cagaagctat tgaagctgag ggagccacag ctcctctgac agagttactt 3121 cactctagga atgaaggtgt ggcgacatat gcagctgctg ttttgttccg aatgtctgag 3181 gacaagccac aagattacaa gaaacggctt tcagttgagc tgaccagctc tctcttcaga 3241 acagagccaa tggcttggaa tgagactgct gatcttggac ttgatattgg tgcccaggga 3301 gaaccccttg gatatcgcca ggatgatcct agctatcgtt cttttcactc tggtggatat 3361 ggccaggatg ccttgggtat ggaccccatg atggaacatg agatgggtgg ccaccaccct 3421 ggtgctgact atccagttga tgggctgcca gatctggggc atgcccagga cctcatggat 3481 gggctgcctc caggtgacag caatcagctg gcctggtttg atactgacct gtaagaccca 3541 gctttcttgt acaaagtggt tgatatccag cacagtggcg gccgctcgag tctagagggc 3601 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc 3661 ggttagtaat gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca 3721 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc 3781 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg 3841 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct 3901 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg 3961 tatccccacg cgccctgtag cggcgcatta aataacttcg tataatgtat gctatacgaa 4021 gttatatcgt atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac 4081 accctaactg acacacattc cacagaattc cttgaaatac atccacaggt acacctccaa 4141 ttgactcaaa tgatgtcaat tagtctatca gaagcttcta aagccatgac atcattttct 4201 ggaattttcc aagctgttta aaggcacagt caacttagtg tatgtaaact tctgacccac 4261 tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa ttgttggaaa 4321 aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac tattgtttgt 4381 taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac ttaagtgtat 4441 gtaaacttcc gacttcaact gtagggtacc gagctcgaat tcactggccg tcgttttaca 4501 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc 4561 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg 4621 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 4681 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 4741 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc 4801 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc 4861 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt 4921 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac 4981 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 5041 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 5101 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 5161 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 5221 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 5281 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 5341 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 5401 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 5461 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 5521 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 5581 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 5641 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 5701 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 5761 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 5821 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 5881 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 5941 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 6001 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 6061 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 6121 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 6181 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 6241 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 6301 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 6361 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 6421 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 6481 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 6541 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 6601 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 6661 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 6721 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 6781 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 6841 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 6901 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 6961 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 7021 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 7081 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag 7141 acctatacag ttgaagtcgg aagtttacat acacttaagt tggagtcatt aaaactcgtt 7201 tttcaactac tccacaaatt tcttgttaac aaacaatagt tttggcaagt cagttaggac 7261 atctactttg tgcatgacac aagtcatttt tccaacaatt gtttacagac agattatttc 7321 acttataatt cactgtatca caattccagt gggtcagaag tttacataca ctaagttgac 7381 tgtgccttta aacagcttgg aaaattccag aaaatgatgt catggcttta gaagcttctg 7441 atagactaat tgacatcatt tgagtcaatt ggaggtgtac ctgtggatgt atttcaaggg 7501 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagcgg 7561 ccgctagctc gagatcttaa ttaaactagt atcgatataa cttcgtataa tgtatgctat 7621 acgaagttat atgtacgggc cagatatacg cgttgacatt gattattgac taggcttttg 7681 c //