LOCUS Exported 5920 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV5-Smad7-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5920) AUTHORS Hayashi H, Abdollah S, Qiu Y, Cai J, Xu YY, Grinnell BW, Richardson MA, Topper JN, Gimbrone MA, Wrana JL, Falb D TITLE The MAD-related protein Smad7 associates with the TGFbeta receptor and functions as an antagonist of TGFbeta signaling. JOURNAL Cell. 1997 Jun 27. 89(7):1165-73. PUBMED 9215638 REFERENCE 2 (bases 1 to 5920) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5920 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 75..97 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 89..106 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 90..106 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 267..646 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 647..850 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 801..821 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 2161..2187 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2210..2832 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2409..2428) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(2856..2876) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2861..3190 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3041..3176 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3103..3122 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3228..3247) /label=T7 /note="T7 promoter, forward primer" promoter complement(3229..3247) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3261..3277) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3261..3277) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3274..3296) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3285..3301 /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(3309..3339) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3354..3375 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3492..3509) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3663..4251) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3743..3762) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4422..5282) /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 5045..5064 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5283..5387) /gene="bla" /label=AmpR promoter rep_origin 5414..5869 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5501..5520) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5711..5732 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt 61 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctgatcta 121 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa 181 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg 241 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc 301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga 541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt 601 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg 661 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc 721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg 781 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat 841 aagcagagct cgtttagtga accgtcagaa ttcgctgctg ctatgttcag gaccaaacga 901 tctgcgctcg tccggcgtct ctggaggagc cgtgcgcccg gcggcgagga cgaggaggag 961 ggcgcagggg gaggtggagg aggaggcgag ctgcggggag aaggggcgac ggacagccga 1021 gcgcatgggg ccggtggcgg cggcccgggc agggctggat gctgcctggg caaggcggtg 1081 cgaggtgcca aaggtcacca ccatccccac ccgccagccg cgggcgccgg cgcggccggg 1141 ggcgccgagg cggatctgaa ggcgctcacg cactcggtgc tcaagaaact gaaggagcgg 1201 cagctggagc tgctgctcca ggccgtggag tcccgcggcg ggacgcgcac cgcgtgcctc 1261 ctgctgcccg gccgcctgga ctgcaggctg ggcccggggg cgcccgccgg cgcgcagcct 1321 gcgcagccgc cctcgtccta ctcgctcccc ctcctgctgt gcaaagtgtt caggtggccg 1381 gatctcaggc attcctcgga agtcaagagg ctgtgttgct gtgaatctta cgggaagatc 1441 aaccccgagc tggtgtgctg caacccccat caccttagcc gactctgcga actagagtct 1501 cccccccctc cttactccag atacccgatg gattttctca aaccaactgc agactgtcca 1561 gatgctgtgc cttcctccgc tgaaacaggg ggaacgaatt atctggcccc tggggggctt 1621 tcagattccc aacttcttct ggagcctggg gatcggtcac actggtgcgt ggtggcatac 1681 tgggaggaga agacgagagt ggggaggctc tactgtgtcc aggagccctc tctggatatc 1741 ttctatgatc tacctcaggg gaatggcttt tgcctcggac agctcaattc ggacaacaag 1801 agtcagctgg tgcagaaggt gcggagcaaa atcggctgcg gcatccagct gacgcgggag 1861 gtggatggtg tgtgggtgta caaccgcagc agttacccca tcttcatcaa gtccgccaca 1921 ctggacaacc cggactccag gacgctgttg gtacacaagg tgttccccgg tttctccatc 1981 aaggctttcg actacgagaa ggcgtacagc ctgcagcggc ccaatgacca cgagtttatg 2041 cagcagccgt ggacgggctt taccgtgcag atcagctttg tgaagggctg gggccagtgc 2101 tacacccgcc agttcatcag cagctgcccg tgctggctag aggtcatctt caacagccgg 2161 tatccgtatg atgttcctga ttatgctagc ctctaatcta gaggatcccg ggtggcatcc 2221 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 2281 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 2341 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 2401 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 2461 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 2521 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 2581 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 2641 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata 2701 actataccag caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt 2761 gggacatttg agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat 2821 gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg 2881 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 2941 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 3001 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 3061 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 3121 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 3181 cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa gttaattccc tatagtgagt 3241 cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 3301 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 3361 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 3421 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 3481 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 3541 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 3601 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 3661 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 3721 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 3781 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 3841 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 3901 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 3961 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 4021 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 4081 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 4141 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 4201 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 4261 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 4321 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 4381 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 4441 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 4501 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 4561 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 4621 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 4681 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 4741 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 4801 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 4861 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 4921 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 4981 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 5041 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 5101 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 5161 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 5221 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 5281 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 5341 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 5401 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 5461 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 5521 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 5581 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 5641 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 5701 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 5761 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 5821 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat 5881 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc //