LOCUS       sequence-197541-        6428 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 6428)
  AUTHORS   Macias-Silva M, Abdollah S, Hoodless PA, Pirone R, Attisano L, Wrana
            JL
  TITLE     MADR2 is a substrate of the TGFbeta receptor and its phosphorylation
            is required for nuclear accumulation and signaling.
  JOURNAL   Cell. 1996 Dec 27. 87(7):1215-24.
  PUBMED    8980228
REFERENCE   2  (bases 1 to 6428)
  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. 1996 
            Dec 27. 87(7):1215-24."
COMMENT     Sequence Label: pCMV5B-Smad4
FEATURES             Location/Qualifiers
     source          1..6428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..154
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     146..163
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     147..163
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        324..703
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        704..907
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     858..878
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2775..3397
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3421..3441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3426..3755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3606..3741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3793..3812)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3794..3812)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3826..3842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3826..3842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3839..3861)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3850..3866
                     /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(3874..3880,3881..3898,3899..3904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3919..3940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4057..4074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4228..4816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4308..4327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4987..5778,5779..5847))
                     /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     5610..5629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5848..5952)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5979..6407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6066..6085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6276..6297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
       61 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      121 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac
      181 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
      241 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
      301 atgtccaata tgaccgccat gttgacattg attattgact agttattaat agtaatcaat
      361 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      421 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      481 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      541 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
      601 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
      661 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      721 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      781 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      841 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      901 cagagctcgt ttagtgaacc gtcagaattg atctggtacc ggatcgaatt caagcaggat
      961 gacctgggct ccgcgccgca ccgcgcctcg ccagatgtct tgacagtttt tcttgcaaca
     1021 ttggccattg gttttcactg ccttcaaaag atcaaaatta ctccagaaat tggagagttg
     1081 gatttaaaag aaaaaacttg aacaaatgga caatatgtct ataacaaata caccaacaag
     1141 taacgatgcc tgtctgagca ttgtacatag tttgatgtgt catagacaag gtggggaaag
     1201 tgaaaccttt gcaaaaagag caattgagag tttggtaaag aagctgaaag agaaaaaaga
     1261 tgaattggat tctttaataa cagctataac tacaaatgga gctcatccta gcaagtgtgt
     1321 caccatacag agaacattgg atggacgact tcaggtggct ggtcggaaag gatttcctca
     1381 tgtgatctat gcccgtctgt ggaggtggcc tgatctacac aagaatgaac taaagcatgt
     1441 taaatattgt cagtatgcgt ttgacttaaa atgtgacagt gtctgtgtga atccatatca
     1501 ctatgagcgg gttgtctcac ctggaattga tctctcagga ttaacactgc agagtaatgt
     1561 tccaagtatg ttagtgaagg atgagtacgt tcacgacttt gaaggacagc cgtccttacc
     1621 cactgaagga cattcgattc aaaccatcca acacccgcca agtaatcgcg catcaacgga
     1681 gacgtacagc gccccggctc tgttagcccc ggcagagtct aacgccacca gcaccaccaa
     1741 cttccccaac attcctgtgg cttccacaag tcagccggcc agtattctgg cgggcagcca
     1801 tagtgaagga ctgttgcaga tagcttcagg gcctcagcca ggacagcagc agaatggatt
     1861 tactgctcag ccagctactt accatcataa cagcactacc acctggactg gaagtaggac
     1921 tgcaccatac acacctaatt tgcctcacca ccaaaacggc catcttcagc accacccgcc
     1981 tatgccgccc catcctggac attactggcc agttcacaat gagcttgcat tccagcctcc
     2041 catttccaat catcctgctc ctgagtactg gtgctccatt gcttactttg aaatggacgt
     2101 tcaggtagga gagacgttta aggtcccttc aagctgccct gttgtgactg tggatggcta
     2161 tgtggatcct tcgggaggag atcgcttttg cttgggtcaa ctctccaatg tccacaggac
     2221 agaagcgatt gagagagcga ggttgcacat aggcaaagga gtgcagttgg aatgtaaagg
     2281 tgaaggtgac gtttgggtca ggtgccttag tgaccacgcg gtctttgtac agagttacta
     2341 cctggacaga gaagctggcc gagcacctgg cgacgctgtt cataagatct acccaagcgc
     2401 gtatataaag gtctttgatc tgcggcagtg tcaccggcag atgcagcaac aggcggccac
     2461 tgcgcaagct gcagctgctg ctcaggcggc ggccgtggca gggaacatcc ctggccctgg
     2521 gtccgtgggt ggaatagctc cagccatcag tctgtctgct gctgctggca tcggtgtgga
     2581 tgacctccgg cgattgtgca ttctcaggat gagctttgtg aagggctggg gcccagacta
     2641 ccccaggcag agcatcaagg aaaccccgtg ctggattgag attcaccttc accgagctct
     2701 gcagctcttg gatgaagtcc tgcacaccat gcccattgcg gacccacagc ctttagactg
     2761 atctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     2821 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     2881 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     2941 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     3001 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     3061 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     3121 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     3181 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     3241 tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag acacagcata
     3301 ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct
     3361 catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt
     3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3661 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3721 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc
     3781 agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc
     3841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     6001 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     6061 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6121 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6181 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6241 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6301 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6361 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6421 aacgctta
//
