LOCUS       sequence-460667-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Comparing with wild type COL1A1 promoter, tested whether mutant TEAD
            binding sites changed the promoter activity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5334)
  AUTHORS   Yang X, Delsante M, Daneshpajouhnejad P, Fenaroli P, Mandell KP, 
            Wang X, Takahashi S, Halushka MK, Kopp JB, Levi M, Rosenberg AZ
  TITLE     Bile Acid Receptor Agonist Reverses Transforming Growth 
            Factor-beta1-Mediated Fibrogenesis in Human Induced Pluripotent Stem
            Cells-Derived Kidney Organoids.
  JOURNAL   Lab Invest. 2024 May;104(5):100336. doi: 
            10.1016/j.labinv.2024.100336. Epub 2024 Jan 22.
  PUBMED    38266922
REFERENCE   2  (bases 1 to 5334)
  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: "Lab 
            Invest."; date: "2024-05"; volume: "104(5):100336. doi"; pages: " 
            10.1016/j.labinv.2024.100336. Epub 2024 Jan 22"
COMMENT     Sequence Label: PGL4.11-COL1A1mTEAD
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1065..1074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1071..2720
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2724..2843
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2895..3016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2906..2925)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2960..2979
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3264..3281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3435..4023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3515..3534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4223..5014,5015..5083))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5188..5236
                     /note="synthetic polyadenylation signal"
     misc_feature    join(5250..5334,1..7)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5290..5309
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 tttctctggc ctaactggcc ggtacctcac caatgatcac aggcctccca ctaaaaatac
       61 ttcccaactc tggggtggaa gagtttgggg gatgaatttt taggggattg caagccccaa
      121 tccccacctc tgtgtcccta gaatccccca cccctacctt ggctgctcca tcacccaacc
      181 accaaagctt tcttctgcag aggccaccta gtcatgtttc tcaccctgca cctcagcctc
      241 cccactccat ctctcaatca tgcctagggt ttggaggaag gcatttgatt ctgttctgga
      301 gcacagcaga agaattgaca tcctcaaaat taaaactccc ttgcctgcac ccctccctca
      361 gatatctgat tcttaatgtc tagaactgcg tctgtaaatt gttccccaaa tattcctaag
      421 ctccatcccc tagccacacc agaagacacc cccaaacagg cacatctttt taattcccag
      481 cttcctctgt tttggagagg tcctcagcat gcctctttat gcccctccct tagctcttgc
      541 caggatatca gagggtgact ggggcacagc caggaggacc ccctccccaa cacccccaac
      601 ccttccacct ttggaagtct ccccacccag ctccccagtt ccccagttcc acttcttcta
      661 gattggaggt cccaggaaga gagcagaggg gcacccctac ccactggtta gcccacgcca
      721 ttctgaggac ccagctgcac ccctaccaca gcacctctgg cccaggctgg gctggggggc
      781 tggggaggca gagctgcgaa gaggggagat gtggggtgga ctcccttccc tcctcctccc
      841 cctctccatt ccaactccca aattgggggc cgggctaggc agctctgatt ggctggggca
      901 cgggcggccg gctccccctc tccgaggggc agggttcctc cctgctctcc atcaggacag
      961 tataaaaggg gcccgggcca gtcgtcggag cagacgggag tttctcgagg atatcaagat
     1021 ctggcctcgg cggccaagct tggcaatccg gtactgttgg taaagccacc atggaagatg
     1081 ccaaaaacat taagaagggc ccagcgccat tctacccact cgaagacggg accgccggcg
     1141 agcagctgca caaagccatg aagcgctacg ccctggtgcc cggcaccatc gcctttaccg
     1201 acgcacatat cgaggtggac attacctacg ccgagtactt cgagatgagc gttcggctgg
     1261 cagaagctat gaagcgctat gggctgaata caaaccatcg gatcgtggtg tgcagcgaga
     1321 atagcttgca gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc
     1381 cagctaacga catctacaac gagcgcgagc tgctgaacag catgggcatc agccagccca
     1441 ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa aagaagctac
     1501 cgatcataca aaagatcatc atcatggata gcaagaccga ctaccagggc ttccaaagca
     1561 tgtacacctt cgtgacttcc catttgccac ccggcttcaa cgagtacgac ttcgtgcccg
     1621 agagcttcga ccgggacaaa accatcgccc tgatcatgaa cagtagtggc agtaccggat
     1681 tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg
     1741 accccatctt cggcaaccag atcatccccg acaccgctat cctcagcgtg gtgccatttc
     1801 accacggctt cggcatgttc accacgctgg gctacttgat ctgcggcttt cgggtcgtgc
     1861 tcatgtaccg cttcgaggag gagctattct tgcgcagctt gcaagactat aagattcaat
     1921 ctgccctgct ggtgcccaca ctatttagct tcttcgctaa gagcactctc atcgacaagt
     1981 acgacctaag caacttgcac gagatcgcca gcggcggggc gccgctcagc aaggaggtag
     2041 gtgaggccgt ggccaaacgc ttccacctac caggcatccg ccagggctac ggcctgacag
     2101 aaacaaccag cgccattctg atcacccccg aaggggacga caagcctggc gcagtaggca
     2161 aggtggtgcc cttcttcgag gctaaggtgg tggacttgga caccggtaag acactgggtg
     2221 tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat catgagcggc tacgttaaca
     2281 accccgaggc tacaaacgct ctcatcgaca aggacggctg gctgcacagc ggcgacatcg
     2341 cctactggga cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaaat
     2401 acaagggcta ccaggtagcc ccagccgaac tggagagcat cctgctgcaa caccccaaca
     2461 tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg cccgccgcag
     2521 tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga gatcgtggac tatgtggcca
     2581 gccaggttac aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta
     2641 aaggactgac cggcaagttg gacgcccgca agatccgcga gattctcatt aaggccaaga
     2701 agggcggcaa gatcgccgtg aattctcacg gcttccctcc cgaggtggag gagcaggccg
     2761 ccggcaccct gcccatgagc tgcgcccagg agagcggcat ggatagacac cctgctgctt
     2821 gcgccagcgc caggatcaac gtctaaggcc gcgactctag agtcggggcg gccggccgct
     2881 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2941 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3001 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3061 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     3121 tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     3181 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     3241 tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3301 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3361 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3421 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3481 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3541 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3601 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3661 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3721 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3781 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3841 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3901 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3961 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4021 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4081 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4141 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc
     4201 aaatgctaaa ccactgcagt ggttaccagt gcttgatcag tgaggcaccg atctcagcga
     4261 tctgcctatt tcgttcgtcc atagtggcct gactccccgt cgtgtagatc actacgattc
     4321 gtgagggctt accatcaggc cccagcgcag caatgatgcc gcgagagccg cgttcaccgg
     4381 cccccgattt gtcagcaatg aaccagccag cagggagggc cgagcgaaga agtggtcctg
     4441 ctactttgtc cgcctccatc cagtctatga gctgctgtcg tgatgctaga gtaagaagtt
     4501 cgccagtgag tagtttccga agagttgtgg ccattgctac tggcatcgtg gtatcacgct
     4561 cgtcgttcgg tatggcttcg ttcaactctg gttcccagcg gtcaagccgg gtcacatgat
     4621 cacccatatt atgaagaaat gcagtcagct ccttagggcc tccgatcgtt gtcagaagta
     4681 agttggccgc ggtgttgtcg ctcatggtaa tggcagcact acacaattct cttaccgtca
     4741 tgccatccgt aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt
     4801 agtgtatacg gcgaccaagc tgctcttgcc cggcgtctat acgggacaac accgcgccac
     4861 atagcagtac tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa
     4921 ggatcttgcc gctattgaga tccagttcga tatagcccac tcttgcaccc agttgatctt
     4981 cagcatcttt tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg
     5041 caaagaaggg aatgagtgcg acacgaaaat gttggatgct catactcgtc ctttttcaat
     5101 attattgaag catttatcag ggttactagt acgtctctca aggataagta agtaatatta
     5161 aggtacggga ggtattggac aggccgcaat aaaatatctt tattttcatt acatctgtgt
     5221 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     5281 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aaca
//
