LOCUS Exported 4542 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CS2 Flag-Smad6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4542) AUTHORS Hata A, Lagna G, Massague J, Hemmati-Brivanlou A TITLE Smad6 inhibits BMP/Smad1 signaling by specifically competing with the Smad4 tumor suppressor. JOURNAL Genes Dev. 1998 Jan 15. 12(2):186-97. PUBMED 9436979 REFERENCE 2 (bases 1 to 4542) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4542 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1..24 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(740..757) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 762..896 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(786..805) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 840..859 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1013..1033) /label=T3 /note="T3 promoter, forward primer" promoter complement(1013..1031) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1052..1068) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1052..1068) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1065..1087) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1076..1092 /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(1100..1130) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1145..1166 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1283..1300) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1454..2042) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1534..1553) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2213..3073) /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 2836..2855 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3074..3178) /gene="bla" /label=AmpR promoter rep_origin complement(3204..3659) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3341..3362) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3553..3572 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gactacaagg acgacgatga caagtccaga atgggcaaac ccatagagac acaaaaatct 61 ccgccacctc cctactctcg gctgtctcct cgcgacgagt acaagccact ggatctgtcc 121 gattccacat tgtcttacac tgaaacggag gctaccaact ccctcatcac tgctccgggt 181 gaattctcag acgccagcat gtctccggac gccaccaagc cgagccactg gtgcagcgtg 241 gcgtactggg agcaccggac gcgcgtgggc cgcctctatg cggtgtacga ccaggccgtc 301 agcatcttct acgacctacc tcagggcagc ggcttctgcc tgggccagct caacctggag 361 cagcgcagcg agtcggtgcg gcgaacgcgc agcaagatcg gcttcggcat cctgctcagc 421 aaggagcccg acggcgtgtg ggcctacaac cgcggcgagc accccatctt cgtcaactcc 481 ccgacgctgg acgcgcccgg cggccgcgcc ctggtcgtgc gcaaggtgcc ccccggctac 541 tccatcaagg tgttcgactt cgagcgctcg ggcctgcagc acgcgcccga gcccgacgcc 601 gccgacggcc cctacgaccc caacagcgtc cgcatcagct tcgccaaggg ctgggggccc 661 tgctactccc ggcagttcat cacctcctgc ccctgctggc tggagatcct cctcaacaac 721 cccagatagt ggtctagaac tatagtgagt cgtattacgt agatccagac atgataagat 781 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg 841 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca 901 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat 961 tcgcggcgcg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt 1021 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 1081 atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg 1141 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 1201 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 1261 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 1321 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 1381 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 1441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 1501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 1561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 1621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 1681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 1741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 1801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 1861 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 1921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 1981 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 2041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 2101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 2161 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 2221 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 2281 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 2341 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 2401 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 2461 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 2521 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 2581 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 2641 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 2701 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 2761 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 2821 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 2881 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 2941 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 3001 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 3061 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 3121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 3181 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt 3241 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta 3301 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc 3361 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg 3421 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact 3481 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt 3541 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc 3601 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc 3661 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 3721 attacgccag tcgatcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa 3781 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa 3841 tatggtggat ctggacccca gccaattcaa tatggctatt ggccaggttc aatactatgt 3901 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc 3961 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc 4021 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc 4081 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc 4141 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg 4201 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg 4261 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc 4321 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc 4381 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg 4441 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg 4501 aaccgccatt ctgcctgggg acgtcggagc aagcttacca tg //