LOCUS       sequence-196957-        5356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SMAD2 (3A) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   Yeh HW, Hsu EC, Lee SS, Lang YD, Lin YC, Chang CY, Lee SY, Gu DL, 
            Shih JH, Ho CM, Chen CF, Chen CT, Tu PH, Cheng CF, Chen RH, Yang RB,
            Jou YS
  TITLE     PSPC1 mediates TGF-beta1 autocrine signalling and Smad2/3 target 
            switching to promote EMT, stemness and metastasis.
  JOURNAL   Nat Cell Biol. 2018 Apr;20(4):479-491. doi: 
            10.1038/s41556-018-0062-y. Epub 2018 Mar 28.
  PUBMED    29593326
REFERENCE   2  (bases 1 to 5356)
  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"; doi: 
            "10.1038/s41556-018-0062-y"; journalName: "Nat Cell Biol"; date: 
            "2018-04"; volume: "20"; issue: "4"; pages: "479-491"
COMMENT     Sequence Label: CS2 Flag-Smad2 (3A)
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             72..95
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1581..1715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1605..1624)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1659..1678
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1832..1852)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1832..1850)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1871..1887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1871..1887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1884..1906)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1895..1911
                     /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(1919..1925,1926..1943,1944..1949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1964..1985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2102..2119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2273..2861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2353..2372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3032..3823,3824..3892))
                     /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     3655..3674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3893..3997)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4023..4478)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4160..4181)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4372..4391
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atttaggtga cactatagaa tacaagctac ttgttctttt tgcaggatcc catcgattcg
       61 aattcaccat ggactacaag gacgacgatg acaagtcgtc catcttgcca ttcacgccgc
      121 cagttgtgaa gagactgctg ggatggaaga agtcagctgg tgggtctgga ggagcaggcg
      181 gaggagagca gaatgggcag gaagaaaagt ggtgtgagaa agcagtgaaa agtctggtga
      241 agaagctaaa gaaaacagga cgattagatg agcttgagaa agccatcacc actcaaaact
      301 gtaatactaa atgtgttacc ataccaagca cttgctctga aatttgggga ctgagtacac
      361 caaatacgat agatcagtgg gatacaacag gcctttacag cttctctgaa caaaccaggt
      421 ctcttgatgg tcgtctccag gtatcccatc gaaaaggatt gccacatgtt atatattgcc
      481 gattatggcg ctggcctgat cttcacagtc atcatgaact caaggcaatt gaaaactgcg
      541 aatatgcttt taatcttaaa aaggatgaag tatgtgtaaa cccttaccac tatcagagag
      601 ttgagacacc agttttgcct ccagtattag tgccccgaca caccgagatc ctaacagaac
      661 ttccgcctct ggatgactat actcactcca ttccagaaaa cactaacttc ccagcaggaa
      721 ttgagccaca gagtaattat attccagaaa cgccacctcc tggatatatc agtgaagatg
      781 gagaaacaag tgaccaacag ttgaatcaaa gtatggacac aggctctcca gcagaactat
      841 ctcctactac tctttcccct gttaatcata gcttggattt acagccagtt acttactcag
      901 aacctgcatt ttggtgttca atagcatatt atgaattaaa tcagagggtt ggagaaacct
      961 tccatgcatc acagccctca ctcactgtag atggctttac agacccatca aattcagaga
     1021 ggttctgctt aggtttactc tccaatgtta accgaaatgc cacggtagaa atgacaagaa
     1081 ggcatatagg aagaggagtg cgcttatact acataggtgg ggaagttttt gctgagtgcc
     1141 taagtgatag tgcaatcttt gtgcagagcc ccaattgtaa tcagagatat ggctggcacc
     1201 ctgcaacagt gtgtaaaatt ccaccaggct gtaatctgaa gatcttcaac aaccaggaat
     1261 ttgctgctct tctggctcag tctgttaatc agggttttga agccgtctat cagctaacta
     1321 gaatgtgcac cataagaatg agttttgtga aagggtgggg agcagaatac cgaaggcaga
     1381 cggtaacaag tactccttgc tggattgaac ttcatctgaa tggacctcta cagtggttgg
     1441 acaaagtatt aactcagatg ggatctcctt cagtgcgttg cgcagcaatg gcataaagct
     1501 tcaccaatca agtcccatga aaagacttaa tgtaacaact cttctcgagc ctctagaact
     1561 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
     1621 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1681 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1741 tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggcgcgc cgcggcgcca
     1801 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     1861 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1921 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1981 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2041 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2101 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2161 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2221 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2281 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2341 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2401 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2461 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2521 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2581 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2641 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2701 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2761 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2821 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2881 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2941 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3001 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3061 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3121 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3181 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3241 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3301 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3361 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3421 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3481 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3541 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3601 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3661 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3721 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3781 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3841 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3901 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3961 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4021 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4081 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4141 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4201 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4261 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4321 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4381 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4441 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     4501 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgatcgaccg
     4561 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     4621 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     4681 ccaattcaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
     4741 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
     4801 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
     4861 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
     4921 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
     4981 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
     5041 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
     5101 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
     5161 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
     5221 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
     5281 gcctaatggg aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga
     5341 cgtcggagca agcttg
//
