LOCUS       sequence-318679-        5385 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 5385)
  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 5385)
  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 HA-Smad6
FEATURES             Location/Qualifiers
     source          1..5385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        657..675
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     657..674
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             713..739
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             719..745
                     /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    2266..2400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2290..2309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2344..2363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2517..2537)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2517..2535)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2556..2572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2556..2572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2569..2591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2580..2596
                     /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(2604..2610,2611..2628,2629..2634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2649..2670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2787..2804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2958..3546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3038..3057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3717..4508,4509..4577))
                     /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     4340..4359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4578..4682)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4708..5163)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4845..4866)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5057..5076
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaggttca atactatgta ttggccctat gccatatagt attccatata tgggttttcc
       61 tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta
      121 ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat
      181 tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac
      241 ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg
      301 tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg
      361 acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca
      421 ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta
      481 aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac
      541 gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat
      601 aagcaatgct cgtttaggga accgccattc tgcctgggga cgtcggagca agcttgattt
      661 aggtgacact atagaataca agctacttgt tctttttgca ggatccacca tgtatccgta
      721 tgatgttcct gattatgcta gcctcttcag gtccaaacgc ttggggctgg tgcggcgact
      781 ttggcgaagt cgtgtggtcc ccgaccggga ggaaggcggc agcggcggcg gcggtggcgg
      841 cgacgaggat gggagcttgg gcagccgagc tgagccggcc ccgcgggcaa gagagggcgg
      901 aggctgcggc cgctccgaag tccgcccggt agccccgcgg cggccccggg acgcagtggg
      961 acagcgaggc gcccagggcg cggggaggcg ccggcgcgca gggggccccc cgaggcccat
     1021 gtcggagcca ggggccggcg ctgggagctc cctgctggac gtggcggagc cgggaggccc
     1081 gggctggctg cccgagagtg actgcgagac ggtgacctgc tgtctctttt cggagcggga
     1141 cgccgccggc gcgccccggg acgccagcga ccccctggcc ggggcggccc tggagccggc
     1201 gggcggcggg cggagtcgcg aagcgcgctc gcggctgctg ctgctggagc aggaactcaa
     1261 aaccgtcacg tactcgctgc tgaagcggct caaggagcgc tcgctggaca cgctgctgga
     1321 ggcggtggag tcccgcggcg gcgtgccggg cggctgcgtg ctggtgccgc gcgccgacct
     1381 ccgcctgggc ggccagcccg cgccgccgca gctgctgctc ggccgcctct ttcgctggcc
     1441 cgacctgcag cacgccgtgg agctgaagcc cctgtgcggc tgccacagct tcgccgccgc
     1501 cgccgacggc cctaccgtgt gctgcaaccc ctaccacttc agccggctct gcgggcccga
     1561 atctccgcca cctccctact ctcggctgtc tcctcgcgac gagtacaagc cactggatct
     1621 gtccgattcc acattgtctt acactgaaac ggaggctacc aactccctca tcactgctcc
     1681 gggtgaattc tcagacgcca gcatgtctcc ggacgccacc aagccgagcc actggtgcag
     1741 cgtggcgtac tgggagcacc ggacgcgcgt gggccgcctc tatgcggtgt acgaccaggc
     1801 cgtcagcatc ttctacgacc tacctcaggg cagcggcttc tgcctgggcc agctcaacct
     1861 ggagcagcgc agcgagtcgg tgcggcgaac gcgcagcaag atcggcttcg gcatcctgct
     1921 cagcaaggag cccgacggcg tgtgggccta caaccgcggc gagcacccca tcttcgtcaa
     1981 ctccccgacg ctggacgcgc ccggcggccg cgccctggtc gtgcgcaagg tgccccccgg
     2041 ctactccatc aaggtgttcg acttcgagcg ctcgggcctg cagcacgcgc ccgagcccga
     2101 cgccgccgac ggcccctacg accccaacag cgtccgcatc agcttcgcca agggctgggg
     2161 gccctgctac tcccggcagt tcatcacctc ctgcccctgc tggctggaga tcctcctcaa
     2221 caaccccaga tagtggtcta gaactatagt gagtcgtatt acgtagatcc agacatgata
     2281 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2341 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2401 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2461 taattcgcgg cgcgccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     2521 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2581 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     2641 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2701 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2761 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3361 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3721 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3781 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3841 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3901 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3961 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4021 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4081 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4141 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4201 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4261 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4321 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4381 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4441 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4501 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4561 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4681 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4741 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4801 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4861 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4921 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4981 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5041 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5101 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5161 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5221 cgctattacg ccagtcgatc gaccgccaat tcaatatggc gtatatggac tcatgccaat
     5281 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
     5341 tcaatatggt ggatctggac cccagccaat tcaatatggc tattg
//
