LOCUS       sequence-460668-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human COL1A1 promoter and testing the promoter 
            activity with luciferase assay..
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-COL1A1WT
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1059..1068
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1065..2714
                     /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             2718..2837
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2889..3010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2900..2919)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2954..2973
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3258..3275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3429..4017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3509..3528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4217..5008,5009..5077))
                     /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    5182..5230
                     /note="synthetic polyadenylation signal"
     misc_feature    join(5244..5334,1)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5284..5303
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 tggcctaact ggccggtacc tcaccaatga tcacaggcct cccactaaaa atacttccca
       61 actctggggt ggaagagttt gggggatgaa tttttagggg attgcaagcc ccaatcccca
      121 cctctgtgtc cctagaatcc cccaccccta ccttggctgc tccatcaccc aaccaccaaa
      181 gctttcttct gcagaggcca cctagtcatg tttctcaccc tgcacctcag cctccccact
      241 ccatctctca atcatgccta gggtttggag gaaggcattt gattctgttc tggagcacag
      301 cagaagaatt gacatcctca aaattaaaac tcccttgcct gcacccctcc ctcagatatc
      361 tgattcttaa tgtctagaaa ggaatctgta aattgttccc caaatattcc taagctccat
      421 cccctagcca caccagaaga cacccccaaa caggcacatc tttttaattc ccagcttcct
      481 ctgttttgga gaggtcctca gcatgcctct ttatgcccct cccttagctc ttgccaggat
      541 atcagagggt gactggggca cagccaggag gaccccctcc ccaacacccc caacccttcc
      601 acctttggaa gtctccccac ccagctcccc agttccccag ttccacttct tctagattgg
      661 aggtcccagg aagagagcag aggggcaccc ctacccactg gttagcccac gccattctga
      721 ggacccagct gcacccctac cacagcacct ctggcccagg ctgggctggg gggctgggga
      781 ggcagagctg cgaagagggg agatgtgggg tggactccct tccctcctcc tccccctctc
      841 cattccaact cccaaattgg gggccgggct aggcagctct gattggctgg ggcacgggcg
      901 gccggctccc cctctccgag gggcagggtt cctccctgct ctccatcagg acagtataaa
      961 aggggcccgg gccagtcgtc ggagcagacg ggagtttctc gaggatatca agatctggcc
     1021 tcggcggcca agcttggcaa tccggtactg ttggtaaagc caccatggaa gatgccaaaa
     1081 acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc
     1141 tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac
     1201 atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag
     1261 ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct
     1321 tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta
     1381 acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg
     1441 tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca
     1501 tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca
     1561 ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct
     1621 tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca
     1681 agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca
     1741 tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg
     1801 gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt
     1861 accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc
     1921 tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc
     1981 taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg
     2041 ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa
     2101 ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg
     2161 tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc
     2221 agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg
     2281 aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact
     2341 gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg
     2401 gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg
     2461 acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg
     2521 tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg
     2581 ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac
     2641 tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg
     2701 gcaagatcgc cgtgaattct cacggcttcc ctcccgaggt ggaggagcag gccgccggca
     2761 ccctgcccat gagctgcgcc caggagagcg gcatggatag acaccctgct gcttgcgcca
     2821 gcgccaggat caacgtctaa ggccgcgact ctagagtcgg ggcggccggc cgcttcgagc
     2881 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2941 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3001 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3061 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     3121 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     3181 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     3241 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3301 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3361 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3601 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3841 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3901 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3961 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4081 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4141 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc
     4201 taaaccactg cagtggttac cagtgcttga tcagtgaggc accgatctca gcgatctgcc
     4261 tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta gatcactacg attcgtgagg
     4321 gcttaccatc aggccccagc gcagcaatga tgccgcgaga gccgcgttca ccggcccccg
     4381 atttgtcagc aatgaaccag ccagcaggga gggccgagcg aagaagtggt cctgctactt
     4441 tgtccgcctc catccagtct atgagctgct gtcgtgatgc tagagtaaga agttcgccag
     4501 tgagtagttt ccgaagagtt gtggccattg ctactggcat cgtggtatca cgctcgtcgt
     4561 tcggtatggc ttcgttcaac tctggttccc agcggtcaag ccgggtcaca tgatcaccca
     4621 tattatgaag aaatgcagtc agctccttag ggcctccgat cgttgtcaga agtaagttgg
     4681 ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa ttctcttacc gtcatgccat
     4741 ccgtaagatg cttttccgtg accggcgagt actcaaccaa gtcgttttgt gagtagtgta
     4801 tacggcgacc aagctgctct tgcccggcgt ctatacggga caacaccgcg ccacatagca
     4861 gtactttgaa agtgctcatc atcgggaatc gttcttcggg gcggaaagac tcaaggatct
     4921 tgccgctatt gagatccagt tcgatatagc ccactcttgc acccagttga tcttcagcat
     4981 cttttacttt caccagcgtt tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga
     5041 agggaatgag tgcgacacga aaatgttgga tgctcatact cgtccttttt caatattatt
     5101 gaagcattta tcagggttac tagtacgtct ctcaaggata agtaagtaat attaaggtac
     5161 gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct gtgtgttggt
     5221 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     5281 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctc
//
