LOCUS       sequence-460669-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Double mutation of TEAD and SMAD binding elements in COL1A1 
            promoter; check 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-COL1A1double mutants
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1046..1055
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1052..2701
                     /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             2705..2824
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2876..2997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2887..2906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2941..2960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3245..3262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3416..4004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3496..3515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4204..4995,4996..5064))
                     /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    5169..5217
                     /note="synthetic polyadenylation signal"
     misc_feature    5231..5322
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5271..5290
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 cggtacctca ccaatgatca caggcctccc actaaaaata cttcccaact ctggggtgga
       61 agagtttggg ggatgaattt ttaggggatt gcaagcccca atccccacct ctgtgtccct
      121 agaatccccc acccctacct tggctgctcc atcacccaac caccaaagct ttcttctgca
      181 gaggccacct agtcatgttt ctcaccctgc acctcagcct ccccactcca tctctcaatc
      241 atgcctaggg tttggaggaa ggcatttgat tctgttctgg agcacagcag aagaattgac
      301 atcctcaaaa ttaaaactcc cttgcctgca cccctccctc agatatctga ttcttaatgg
      361 accgaactgc gtctgtaaat tgttccccaa atattcctaa gctccatccc ctagccacac
      421 cagaagacac ccccaaacag gcacatcttt ttaattccca gcttcctctg ttttggagag
      481 gtcctcagca tgcctcttta tgcccctccc ttagctcttg ccaggatatc agagggtgac
      541 tggggcacag ccaggaggac cccctcccca acacccccaa cccttccacc tttggaagtc
      601 tccccaccca gctccccagt tccccagttc cacttcttct agattggagg tcccaggaag
      661 agagcagagg ggcaccccta cccactggtt agcccacgcc attctgagga cccagctgca
      721 cccctaccac agcacctctg gcccaggctg ggctgggggg ctggggaggc agagctgcga
      781 agaggggaga tgtggggtgg actcccttcc ctcctcctcc ccctctccat tccaactccc
      841 aaattggggg ccgggctagg cagctctgat tggctggggc acgggcggcc ggctccccct
      901 ctccgagggg cagggttcct ccctgctctc catcaggaca gtataaaagg ggcccgggcc
      961 agtcgtcgga gcagacggga gtttctcgag gatatcaaga tctggcctcg gcggccaagc
     1021 ttggcaatcc ggtactgttg gtaaagccac catggaagat gccaaaaaca ttaagaaggg
     1081 cccagcgcca ttctacccac tcgaagacgg gaccgccggc gagcagctgc acaaagccat
     1141 gaagcgctac gccctggtgc ccggcaccat cgcctttacc gacgcacata tcgaggtgga
     1201 cattacctac gccgagtact tcgagatgag cgttcggctg gcagaagcta tgaagcgcta
     1261 tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag aatagcttgc agttcttcat
     1321 gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc ccagctaacg acatctacaa
     1381 cgagcgcgag ctgctgaaca gcatgggcat cagccagccc accgtcgtat tcgtgagcaa
     1441 gaaagggctg caaaagatcc tcaacgtgca aaagaagcta ccgatcatac aaaagatcat
     1501 catcatggat agcaagaccg actaccaggg cttccaaagc atgtacacct tcgtgacttc
     1561 ccatttgcca cccggcttca acgagtacga cttcgtgccc gagagcttcg accgggacaa
     1621 aaccatcgcc ctgatcatga acagtagtgg cagtaccgga ttgcccaagg gcgtagccct
     1681 accgcaccgc accgcttgtg tccgattcag tcatgcccgc gaccccatct tcggcaacca
     1741 gatcatcccc gacaccgcta tcctcagcgt ggtgccattt caccacggct tcggcatgtt
     1801 caccacgctg ggctacttga tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga
     1861 ggagctattc ttgcgcagct tgcaagacta taagattcaa tctgccctgc tggtgcccac
     1921 actatttagc ttcttcgcta agagcactct catcgacaag tacgacctaa gcaacttgca
     1981 cgagatcgcc agcggcgggg cgccgctcag caaggaggta ggtgaggccg tggccaaacg
     2041 cttccaccta ccaggcatcc gccagggcta cggcctgaca gaaacaacca gcgccattct
     2101 gatcaccccc gaaggggacg acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga
     2161 ggctaaggtg gtggacttgg acaccggtaa gacactgggt gtgaaccagc gcggcgagct
     2221 gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc
     2281 tctcatcgac aaggacggct ggctgcacag cggcgacatc gcctactggg acgaggacga
     2341 gcacttcttc atcgtggacc ggctgaagag cctgatcaaa tacaagggct accaggtagc
     2401 cccagccgaa ctggagagca tcctgctgca acaccccaac atcttcgacg ccggggtcgc
     2461 cggcctgccc gacgacgatg ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg
     2521 taaaaccatg accgagaagg agatcgtgga ctatgtggcc agccaggtta caaccgccaa
     2581 gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt
     2641 ggacgcccgc aagatccgcg agattctcat taaggccaag aagggcggca agatcgccgt
     2701 gaattctcac ggcttccctc ccgaggtgga ggagcaggcc gccggcaccc tgcccatgag
     2761 ctgcgcccag gagagcggca tggatagaca ccctgctgct tgcgccagcg ccaggatcaa
     2821 cgtctaaggc cgcgactcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
     2881 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2941 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3001 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3061 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtcgac cgatgccctt
     3121 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     3181 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
     3241 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3301 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3361 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3421 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3481 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3541 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3601 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3661 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3721 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3781 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3841 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3901 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3961 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4021 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4081 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4141 tctaaagtat atatgagtaa acttggtctg acagcggccg caaatgctaa accactgcag
     4201 tggttaccag tgcttgatca gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc
     4261 catagtggcc tgactccccg tcgtgtagat cactacgatt cgtgagggct taccatcagg
     4321 ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat
     4381 gaaccagcca gcagggaggg ccgagcgaag aagtggtcct gctactttgt ccgcctccat
     4441 ccagtctatg agctgctgtc gtgatgctag agtaagaagt tcgccagtga gtagtttccg
     4501 aagagttgtg gccattgcta ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc
     4561 gttcaactct ggttcccagc ggtcaagccg ggtcacatga tcacccatat tatgaagaaa
     4621 tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc
     4681 gctcatggta atggcagcac tacacaattc tcttaccgtc atgccatccg taagatgctt
     4741 ttccgtgacc ggcgagtact caaccaagtc gttttgtgag tagtgtatac ggcgaccaag
     4801 ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca catagcagta ctttgaaagt
     4861 gctcatcatc gggaatcgtt cttcggggcg gaaagactca aggatcttgc cgctattgag
     4921 atccagttcg atatagccca ctcttgcacc cagttgatct tcagcatctt ttactttcac
     4981 cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc
     5041 gacacgaaaa tgttggatgc tcatactcgt cctttttcaa tattattgaa gcatttatca
     5101 gggttactag tacgtctctc aaggataagt aagtaatatt aaggtacggg aggtattgga
     5161 caggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     5221 cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat
     5281 aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctggcctaac tggc
//
