LOCUS       sequence-321809-        4542 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 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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes Dev. 
            1998 Jan 15. 12(2):186-97."
COMMENT     Sequence Label: CS2 Flag-Smad6
FEATURES             Location/Qualifiers
     source          1..4542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..24
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     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(join(1100..1106,1107..1124,1125..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(join(2213..3004,3005..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
//
