LOCUS Exported 5356 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CS2 Flag-Smad2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5356) AUTHORS Hata A, Lo RS, Wotton D, Lagna G, Massague J TITLE Mutations increasing autoinhibition inactivate tumour suppressors Smad2 and Smad4. JOURNAL Nature. 1997 Jul 3. 388(6637):82-7. PUBMED 9214507 REFERENCE 2 (bases 1 to 5356) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5356 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 96..114 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 96..113 /label=SP6 /note="SP6 promoter, forward primer" CDS 167..190 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(1654..1671) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 1676..1810 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1700..1719) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1754..1773 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1927..1947) /label=T3 /note="T3 promoter, forward primer" promoter complement(1927..1945) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1966..1982) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1966..1982) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1979..2001) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1990..2006 /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(2014..2044) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2059..2080 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2197..2214) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2368..2956) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2448..2467) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3127..3987) /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 3750..3769 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3988..4092) /gene="bla" /label=AmpR promoter rep_origin complement(4118..4573) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4255..4276) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4467..4486 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa 61 ccgccattct gcctggggac gtcggagcaa gcttgattta ggtgacacta tagaatacaa 121 gctacttgtt ctttttgcag gatcccatcg attcgaattc accatggact acaaggacga 181 cgatgacaag tcgtccatct tgccattcac gccgccagtt gtgaagagac tgctgggatg 241 gaagaagtca gctggtgggt ctggaggagc aggcggagga gagcagaatg ggcaggaaga 301 aaagtggtgt gagaaagcag tgaaaagtct ggtgaagaag ctaaagaaaa caggacgatt 361 agatgagctt gagaaagcca tcaccactca aaactgtaat actaaatgtg ttaccatacc 421 aagcacttgc tctgaaattt ggggactgag tacaccaaat acgatagatc agtgggatac 481 aacaggcctt tacagcttct ctgaacaaac caggtctctt gatggtcgtc tccaggtatc 541 ccatcgaaaa ggattgccac atgttatata ttgccgatta tggcgctggc ctgatcttca 601 cagtcatcat gaactcaagg caattgaaaa ctgcgaatat gcttttaatc ttaaaaagga 661 tgaagtatgt gtaaaccctt accactatca gagagttgag acaccagttt tgcctccagt 721 attagtgccc cgacacaccg agatcctaac agaacttccg cctctggatg actatactca 781 ctccattcca gaaaacacta acttcccagc aggaattgag ccacagagta attatattcc 841 agaaacgcca cctcctggat atatcagtga agatggagaa acaagtgacc aacagttgaa 901 tcaaagtatg gacacaggct ctccagcaga actatctcct actactcttt cccctgttaa 961 tcatagcttg gatttacagc cagttactta ctcagaacct gcattttggt gttcaatagc 1021 atattatgaa ttaaatcaga gggttggaga aaccttccat gcatcacagc cctcactcac 1081 tgtagatggc tttacagacc catcaaattc agagaggttc tgcttaggtt tactctccaa 1141 tgttaaccga aatgccacgg tagaaatgac aagaaggcat ataggaagag gagtgcgctt 1201 atactacata ggtggggaag tttttgctga gtgcctaagt gatagtgcaa tctttgtgca 1261 gagccccaat tgtaatcaga gatatggctg gcaccctgca acagtgtgta aaattccacc 1321 aggctgtaat ctgaagatct tcaacaacca ggaatttgct gctcttctgg ctcagtctgt 1381 taatcagggt tttgaagccg tctatcagct aactagaatg tgcaccataa gaatgagttt 1441 tgtgaaaggg tggggagcag aataccgaag gcagacggta acaagtactc cttgctggat 1501 tgaacttcat ctgaatggac ctctacagtg gttggacaaa gtattaactc agatgggatc 1561 tccttcagtg cgttgctcaa gcatgtcata aagcttcacc aatcaagtcc catgaaaaga 1621 cttaatgtaa caactcttct cgagcctcta gaactatagt gagtcgtatt acgtagatcc 1681 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa 1741 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa 1801 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg 1861 ggaggttttt taattcgcgg cgcgccgcgg cgccaatgca ttgggcccgg tacccagctt 1921 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc 1981 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg 2041 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 2101 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 2161 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 2221 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 2281 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 2341 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 2401 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 2461 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 2521 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 2581 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 2641 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 2701 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 2761 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 2821 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 2881 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 2941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 3001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 3061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 3121 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 3181 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 3241 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 3301 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 3361 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 3421 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 3481 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 3541 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 3601 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 3661 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 3721 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 3781 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 3841 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 3901 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 3961 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 4021 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 4081 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg 4141 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc 4201 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt 4261 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc 4321 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg 4381 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga 4441 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg 4501 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg 4561 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 4621 cgggcctctt cgctattacg ccagtcgatc gaccgccaat tcaatatggc gtatatggac 4681 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac 4741 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc tattggccag 4801 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg 4861 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc 4921 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg 4981 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa 5041 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat 5101 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt 5161 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact 5221 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg 5281 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa 5341 tgggcgttcc attgac //