LOCUS       sequence-318677-        5352 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 5352)
  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 5352)
  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 Smad6
FEATURES             Location/Qualifiers
     source          1..5352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        658..676
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     658..675
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    2234..2368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2258..2277)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2312..2331
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2485..2505)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2485..2503)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2524..2540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2524..2540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2537..2559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2548..2564
                     /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(2572..2578,2579..2596,2597..2602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2617..2638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2755..2772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2926..3514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3006..3025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3685..4476,4477..4545))
                     /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     4308..4327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4546..4650)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4676..5131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4813..4834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5025..5044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
       61 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
      121 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
      181 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
      241 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
      301 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
      361 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
      421 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
      481 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
      541 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
      601 taagcaatgc tcgtttaggg aaccgccatt ctgcctgggg acgtcggagc aagcttgatt
      661 taggtgacac tatagaatac aagctacttg ttctttttgc aggatccggt atgttcaggt
      721 ccaaacgctc ggggctggtg cggcgacttt ggcgaagtcg tgtggtcccc gaccgggagg
      781 aaggcggcag cggcggcggc ggtggcggcg acgaggatgg gagcttgggc agccgagctg
      841 agccggcccc gcgggcaaga gagggcggag gctgcggccg ctccgaagtc cgcccggtag
      901 ccccgcggcg gccccgggac gcagtgggac agcgaggcgc ccagggcgcg gggaggcgcc
      961 ggcgcgcagg gggccccccg aggcccatgt cggagccagg ggccggcgct gggagctccc
     1021 tgctggacgt ggcggagccg ggaggcccgg gctggctgcc cgagagtgac tgcgagacgg
     1081 tgacctgctg tctcttttcg gagcgggacg ccgccggcgc gccccgggac gccagcgacc
     1141 ccctggccgg ggcggccctg gagccggcgg gcggcgggcg gagtcgcgaa gcgcgctcgc
     1201 ggctgctgct gctggagcag gaactcaaaa ccgtcacgta ctcgctgctg aagcggctca
     1261 aggagcgctc gctggacacg ctgctggagg cggtggagtc ccgcggcggc gtgccgggcg
     1321 gctgcgtgct ggtgccgcgc gccgacctcc gcctgggcgg ccagcccgcg ccgccgcagc
     1381 tgctgctcgg ccgcctcttt cgctggcccg acctgcagca cgccgtggag ctgaagcccc
     1441 tgtgcggctg ccacagcttc gccgccgccg ccgacggccc taccgtgtgc tgcaacccct
     1501 accacttcag ccggctctgc gggcccgaat ctccgccacc tccctactct cggctgtctc
     1561 ctcgcgacga gtacaagcca ctggatctgt ccgattccac attgtcttac actgaaacgg
     1621 aggctaccaa ctccctcatc actgctccgg gtgaattctc agacgccagc atgtctccgg
     1681 acgccaccaa gccgagccac tggtgcagcg tggcgtactg ggagcaccgg acgcgcgtgg
     1741 gccgcctcta tgcggtgtac gaccaggccg tcagcatctt ctacgaccta cctcagggca
     1801 gcggcttctg cctgggccag ctcaacctgg agcagcgcag cgagtcggtg cggcgaacgc
     1861 gcagcaagat cggcttcggc atcctgctca gcaaggagcc cgacggcgtg tgggcctaca
     1921 accgcggcga gcaccccatc ttcgtcaact ccccgacgct ggacgcgccc ggcggccgcg
     1981 ccctggtcgt gcgcaaggtg ccccccggct actccatcaa ggtgttcgac ttcgagcgct
     2041 cgggcctgca gcacgcgccc gagcccgacg ccgccgacgg cccctacgac cccaacagcg
     2101 tccgcatcag cttcgccaag ggctgggggc cctgctactc ccggcagttc atcacctcct
     2161 gcccctgctg gctggagatc ctcctcaaca accccagata gtggtctaga actatagtga
     2221 gtcgtattac gtagatccag acatgataag atacattgat gagtttggac aaaccacaac
     2281 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2341 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2401 ggttcagggg gaggtgtggg aggtttttta attcgcggcg cgccgcggcg ccaatgcatt
     2461 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2521 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2581 agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2641 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2701 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3361 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4681 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4741 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4801 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4861 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4921 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4981 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5041 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5101 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     5161 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgatcga ccgccaattc
     5221 aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct gtgccaattc
     5281 aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc cagccaattc
     5341 aatatggcta tt
//
