LOCUS Exported 6082 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV5 Smad1-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6082) AUTHORS Liu F, Hata A, Baker JC, Doody J, Carcamo J, Harland RM, Massague J TITLE A human Mad protein acting as a BMP-regulated transcriptional activator. JOURNAL Nature. 1996 Jun 13. 381(6583):620-3. PUBMED 8637600 REFERENCE 2 (bases 1 to 6082) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6082 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 82..104 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 96..113 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 97..113 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 274..653 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 654..857 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 808..828 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 2336..2362 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2379..3001 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2578..2597) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(3025..3045) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3030..3359 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3210..3345 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3272..3291 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3397..3416) /label=T7 /note="T7 promoter, forward primer" promoter complement(3398..3416) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3430..3446) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3430..3446) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3443..3465) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3454..3470 /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(3478..3508) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3523..3544 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3661..3678) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3832..4420) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3912..3931) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4591..5451) /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 5214..5233 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5452..5556) /gene="bla" /label=AmpR promoter rep_origin 5583..6038 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5670..5689) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5880..5901 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 61 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 121 tgatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat 181 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt 241 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat 301 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 361 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 421 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 481 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc 541 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac 601 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 661 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 721 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa 781 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg 841 tctatataag cagagctcgt ttagtgaacc gtcagaattc agatctggta ccacgcgtat 901 cgataagctt tatccaagga gtataactag tgctgtcatt atgaatgtga caagtttatt 961 ttcctttaca agtccagctg tgaagagact tcttgggtgg aaacagggcg atgaagaaga 1021 aaaatgggca gagaaagctg ttgatgcttt ggtgaaaaaa ctgaagaaaa agaaaggtgc 1081 catggaggaa ctggaaaagg ccttgagctg cccagggcaa ccgagtaact gtgtcaccat 1141 tccccgctct ctggatggca ggctgcaagt ctcccaccgg aagggactgc ctcatgtcat 1201 ttactgccgt gtgtggcgct ggcccgatct tcagagccac catgaactaa aaccactgga 1261 atgctgtgag tttccttttg gttccaagca gaaggaggtc tgcatcaatc cctaccacta 1321 taagagagta gaaagccctg tacttcctcc tgtgctggtt ccaagacaca gcgaatataa 1381 tcctcagcac agcctcttag ctcagttccg taacttagga caaaatgagc ctcacatgcc 1441 actcaacgcc acttttccag attctttcca gcaacccaac agccacccgt ttcctcactc 1501 tcccaatagc agttacccaa actctcctgg gagcagcagc agcacctacc ctcactctcc 1561 caccagctca gacccaggaa gccctttcca gatgccagct gatacgcccc cacctgctta 1621 cctgcctcct gaagacccca tgacccagga tggctctcag ccgatggaca caaacatgat 1681 ggcgcctccc ctgccctcag aaatcaacag aggagatgtt caggcggttg cttatgagga 1741 accaaaacac tggtgctcta ttgtctacta tgagctcaac aatcgtgtgg gtgaagcgtt 1801 ccatgcctcc tccacaagtg tgttggtgga tggtttcact gatccttcca acaataagaa 1861 ccgtttctgc cttgggctgc tctccaatgt taaccggaat tccactattg aaaacaccag 1921 gcggcatatt ggaaaaggag ttcatcttta ttatgttgga ggggaggtgt atgccgaatg 1981 ccttagtgac agtagcatct ttgtgcaaag tcggaactgc aactaccatc atggatttca 2041 tcctactact gtttgcaaga tccctagtgg gtgtagtctg aaaattttta acaaccaaga 2101 atttgctcag ttattggcac agtctgtgaa ccatggattt gagacagtct atgagcttac 2161 aaaaatgtgt actatacgta tgagctttgt gaagggctgg ggagcagaat accaccgcca 2221 ggatgttact agcaccccct gctggattga gatacatctg cacggccccc tccagtggct 2281 ggataaagtt cttactcaaa tgggttcacc tcataatcct atttcatctg tatcttatcc 2341 gtatgatgtt cctgattatg ctagtctcta gggatcccgg gtggcatccc tgtgacccct 2401 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa 2461 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga 2521 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct 2581 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct 2641 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga 2701 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc 2761 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat 2821 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc 2881 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga 2941 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat 3001 tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3181 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3241 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 3301 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3361 agctcctcga ggaactgaaa aaccagaaag ttaattccct atagtgagtc gtattaaatt 3421 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 3481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 3541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 3601 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 3661 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 3721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 3781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 3841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 3901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 3961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 4261 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 4321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 4381 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 4441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 4501 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 4561 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 4621 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 4681 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 4741 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 4801 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 4861 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 4921 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 4981 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 5041 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 5101 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 5161 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 5221 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 5281 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 5341 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 5401 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 5461 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 5521 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 5581 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 5641 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 5701 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 5761 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 5821 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 5881 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 5941 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 6001 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc 6061 aactgttggg aagggcgatc gg //