LOCUS       sequence-229773-        5920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1997 
            Jun 27. 89(7):1165-73."
COMMENT     Sequence Label: pCMV5-Smad7-HA
FEATURES             Location/Qualifiers
     source          1..5920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             2167..2193
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     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     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(join(3309..3315,3316..3333,3334..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(join(4422..5213,5214..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
//
