LOCUS       sequence-196956-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TGFb1 promoter reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5615)
  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 5615)
  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: pGL3-TGFb1
FEATURES             Location/Qualifiers
     source          1..5615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      886..895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             892..2544
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(958..976)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2585..2706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2596..2615)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2650..2669
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2954..2971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3125..3713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3205..3224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3884..4675,4676..4744))
                     /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     4507..4526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4745..4849)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4876..5331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4963..4982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5173..5194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5462..5510
                     /note="synthetic polyadenylation signal"
     misc_feature    5524..5615
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5564..5583
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atcgataggt acctgggcag ttggcgagaa cagttggcac gggctttcgt gggtggtggg
       61 ccgcagctgc tgcatgggga caccatctac agtggggccg accgctatcg cctgcacaca
      121 gctgctggtg gcaccgtgca cctggagatc ggcctgctgc tccgcaactt cgaccgctac
      181 ggcgtggagt gctgagggac tctgcctcca acgtcaccac catccacacc ccggacaccc
      241 agtgatgggg gaggatggca cagtggtcaa gagcacagac tctagagact gtcagagctg
      301 accccagcta aggcatggca ccgcttctgt cctttctagg acctcggggt ccctctgggc
      361 ccagtttccc tatctgtaaa ttggggacag taaatgtatg gggtcgcagg gtgttgagtg
      421 acaggaggct gcttagccac atgggaggtg ctcagtaaag gagagcaatt cttacaggtg
      481 tctgcctcct gacccttcca tccctcaggt gtcctgttgc cccctcctcc cactgacacc
      541 ctccggaggc ccccatgttg acagaccctc ttctcctacc ttgtttccca gcctgactct
      601 ccttccgttc tgggtccccc tcctctggtc ggctcccctg tgtctcatcc cccggattaa
      661 gccttctccg cctggtcctc tttctctggt gacccacacc gcccgcaaag ccacagcgca
      721 tctggatcac ccgctttggt ggcgcttggc cgccaggagg cagcaccctg tttgcggggc
      781 ggagccgggg tgcccgcccc ctttccccca gggctgaagg gacccccctc ggagcccgcc
      841 cacgcgagat gaggacaagc ttggcattcc ggtactgttg gtaaagccac catggaagac
      901 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
      961 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
     1021 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     1081 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     1141 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     1201 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     1261 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     1321 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     1381 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     1441 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
     1501 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     1561 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     1621 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     1681 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     1741 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     1801 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     1861 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     1921 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     1981 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     2041 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     2101 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     2161 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     2221 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     2281 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     2341 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     2401 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     2461 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     2521 aagggcggaa agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac
     2581 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2641 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2701 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2761 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     2821 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     2881 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     2941 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3001 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3061 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3901 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3961 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4021 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4081 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4141 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4201 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4261 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4321 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4381 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4441 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4501 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4561 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4621 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4681 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4741 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4801 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4861 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4921 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4981 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5041 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5101 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5161 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5221 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5281 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc
     5341 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5401 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     5461 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     5521 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     5581 ccccagtgca agtgcaggtg ccagaacatt tctct
//
