LOCUS       sequence-458138-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tested the COL1A1 promoter activity after abolition of the proximal 
            SMAD binding sites in the promoter..
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-COL1A1mSMAD
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1096..1105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1102..2751
                     /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             2755..2874
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2926..3047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2937..2956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2991..3010
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3295..3312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3466..4054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3546..3565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4254..5045,5046..5114))
                     /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    5219..5267
                     /note="synthetic polyadenylation signal"
     misc_feature    join(5281..5334,1..38)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     join(5321..5334,1..6)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 tgtccccagt gcaagtgcag gtgccagaac atttctctgg cctaactggc cggtacctca
       61 ccaatgatca caggcctccc actaaaaata cttcccaact ctggggtgga agagtttggg
      121 ggatgaattt ttaggggatt gcaagcccca atccccacct ctgtgtccct agaatccccc
      181 acccctacct tggctgctcc atcacccaac caccaaagct ttcttctgca gaggccacct
      241 agtcatgttt ctcaccctgc acctcagcct ccccactcca tctctcaatc atgcctaggg
      301 tttggaggaa ggcatttgat tctgttctgg agcacagcag aagaattgac atcctcaaaa
      361 ttaaaactcc cttgcctgca cccctccctc agatatctga ttcttaatgg accgaaagga
      421 atctgtaaat tgttccccaa atattcctaa gctccatccc ctagccacac cagaagacac
      481 ccccaaacag gcacatcttt ttaattccca gcttcctctg ttttggagag gtcctcagca
      541 tgcctcttta tgcccctccc ttagctcttg ccaggatatc agagggtgac tggggcacag
      601 ccaggaggac cccctcccca acacccccaa cccttccacc tttggaagtc tccccaccca
      661 gctccccagt tccccagttc cacttcttct agattggagg tcccaggaag agagcagagg
      721 ggcaccccta cccactggtt agcccacgcc attctgagga cccagctgca cccctaccac
      781 agcacctctg gcccaggctg ggctgggggg ctggggaggc agagctgcga agaggggaga
      841 tgtggggtgg actcccttcc ctcctcctcc ccctctccat tccaactccc aaattggggg
      901 ccgggctagg cagctctgat tggctggggc acgggcggcc ggctccccct ctccgagggg
      961 cagggttcct ccctgctctc catcaggaca gtataaaagg ggcccgggcc agtcgccgga
     1021 gcagacggga gtttctcgag gatatcaaga tctggcctcg gcggccaagc ttggcaatcc
     1081 ggtactgttg gtaaagccac catggaagat gccaaaaaca ttaagaaggg cccagcgcca
     1141 ttctacccac tcgaagacgg gaccgccggc gagcagctgc acaaagccat gaagcgctac
     1201 gccctggtgc ccggcaccat cgcctttacc gacgcacata tcgaggtgga cattacctac
     1261 gccgagtact tcgagatgag cgttcggctg gcagaagcta tgaagcgcta tgggctgaat
     1321 acaaaccatc ggatcgtggt gtgcagcgag aatagcttgc agttcttcat gcccgtgttg
     1381 ggtgccctgt tcatcggtgt ggctgtggcc ccagctaacg acatctacaa cgagcgcgag
     1441 ctgctgaaca gcatgggcat cagccagccc accgtcgtat tcgtgagcaa gaaagggctg
     1501 caaaagatcc tcaacgtgca aaagaagcta ccgatcatac aaaagatcat catcatggat
     1561 agcaagaccg actaccaggg cttccaaagc atgtacacct tcgtgacttc ccatttgcca
     1621 cccggcttca acgagtacga cttcgtgccc gagagcttcg accgggacaa aaccatcgcc
     1681 ctgatcatga acagtagtgg cagtaccgga ttgcccaagg gcgtagccct accgcaccgc
     1741 accgcttgtg tccgattcag tcatgcccgc gaccccatct tcggcaacca gatcatcccc
     1801 gacaccgcta tcctcagcgt ggtgccattt caccacggct tcggcatgtt caccacgctg
     1861 ggctacttga tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc
     1921 ttgcgcagct tgcaagacta taagattcaa tctgccctgc tggtgcccac actatttagc
     1981 ttcttcgcta agagcactct catcgacaag tacgacctaa gcaacttgca cgagatcgcc
     2041 agcggcgggg cgccgctcag caaggaggta ggtgaggccg tggccaaacg cttccaccta
     2101 ccaggcatcc gccagggcta cggcctgaca gaaacaacca gcgccattct gatcaccccc
     2161 gaaggggacg acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg
     2221 gtggacttgg acaccggtaa gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt
     2281 ggccccatga tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc tctcatcgac
     2341 aaggacggct ggctgcacag cggcgacatc gcctactggg acgaggacga gcacttcttc
     2401 atcgtggacc ggctgaagag cctgatcaaa tacaagggct accaggtagc cccagccgaa
     2461 ctggagagca tcctgctgca acaccccaac atcttcgacg ccggggtcgc cggcctgccc
     2521 gacgacgatg ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg
     2581 accgagaagg agatcgtgga ctatgtggcc agccaggtta caaccgccaa gaagctgcgc
     2641 ggtggtgttg tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc
     2701 aagatccgcg agattctcat taaggccaag aagggcggca agatcgccgt gaattctcac
     2761 ggcttccctc ccgaggtgga ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag
     2821 gagagcggca tggatagaca ccctgctgct tgcgccagcg ccaggatcaa cgtctaaggc
     2881 cgcgactcta gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg
     2941 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3001 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3061 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3121 acctctacaa atgtggtaaa atcgataagg atccgtcgac cgatgccctt gagagccttc
     3181 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     3241 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     3301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4201 atatgagtaa acttggtctg acagcggccg caaatgctaa accactgcag tggttaccag
     4261 tgcttgatca gtgaggcacc gatctcagcg atctgcctat ttcgttcgtc catagtggcc
     4321 tgactccccg tcgtgtagat cactacgatt cgtgagggct taccatcagg ccccagcgca
     4381 gcaatgatgc cgcgagagcc gcgttcaccg gcccccgatt tgtcagcaat gaaccagcca
     4441 gcagggaggg ccgagcgaag aagtggtcct gctactttgt ccgcctccat ccagtctatg
     4501 agctgctgtc gtgatgctag agtaagaagt tcgccagtga gtagtttccg aagagttgtg
     4561 gccattgcta ctggcatcgt ggtatcacgc tcgtcgttcg gtatggcttc gttcaactct
     4621 ggttcccagc ggtcaagccg ggtcacatga tcacccatat tatgaagaaa tgcagtcagc
     4681 tccttagggc ctccgatcgt tgtcagaagt aagttggccg cggtgttgtc gctcatggta
     4741 atggcagcac tacacaattc tcttaccgtc atgccatccg taagatgctt ttccgtgacc
     4801 ggcgagtact caaccaagtc gttttgtgag tagtgtatac ggcgaccaag ctgctcttgc
     4861 ccggcgtcta tacgggacaa caccgcgcca catagcagta ctttgaaagt gctcatcatc
     4921 gggaatcgtt cttcggggcg gaaagactca aggatcttgc cgctattgag atccagttcg
     4981 atatagccca ctcttgcacc cagttgatct tcagcatctt ttactttcac cagcgtttcg
     5041 gggtgtgcaa aaacaggcaa gcaaaatgcc gcaaagaagg gaatgagtgc gacacgaaaa
     5101 tgttggatgc tcatactcgt cctttttcaa tattattgaa gcatttatca gggttactag
     5161 tacgtctctc aaggataagt aagtaatatt aaggtacggg aggtattgga caggccgcaa
     5221 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     5281 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggc
//
