LOCUS       sequence-383810-        4770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prg4 enhancer/promoterfirefly luciferase constructs were constructed
            by cloning various Prg4 regulatory fragments into pGL4.10[luc2] 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4770)
  AUTHORS   Zhang CH, Gao Y, Jadhav U, Hung HH, Holton KM, Grodzinsky AJ, 
            Shivdasani RA, Lassar AB
  TITLE     Creb5 establishes the competence for Prg4 expression in articular 
            cartilage.
  JOURNAL   Commun Biol. 2021 Mar 12;4(1):332. doi: 10.1038/s42003-021-01857-0.
  PUBMED    33712729
REFERENCE   2  (bases 1 to 4770)
  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: "Commun 
            Biol."; date: "2021-03-12"; volume: "4(1):332. doi"; pages: " 
            10.1038/s42003-021-01857-0"
COMMENT     Sequence Label: GL4.10 Prg4 promoter-luciferase
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(322..1113,1114..1182))
                     /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    1287..1335
                     /note="synthetic polyadenylation signal"
     misc_feature    1349..1440
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1389..1408
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2062..2071
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2068..3720
                     /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"
     polyA_signal    3764..3885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3775..3794)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3829..3848
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4133..4150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4304..4770,1..122))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4384..4403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
       61 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      181 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agcggccgca
      301 aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga tctcagcgat
      361 ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca ctacgattcg
      421 tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc gttcaccggc
      481 ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa gtggtcctgc
      541 tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag taagaagttc
      601 gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg tatcacgctc
      661 gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg tcacatgatc
      721 acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg tcagaagtaa
      781 gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc ttaccgtcat
      841 gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt tttgtgagta
      901 gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca ccgcgccaca
      961 tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga aagactcaag
     1021 gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca gttgatcttc
     1081 agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc
     1141 aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc tttttcaata
     1201 ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa gtaatattaa
     1261 ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta catctgtgtg
     1321 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
     1381 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
     1441 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaagatc ttaaaggtgc
     1501 ttgaaaagat ttataaggat agtagttgga gagtttagaa aagaatcaca tttatgtaat
     1561 acacctgccg gaggcttatg aatttatatt gagagttaag agaatgttgg aatgataact
     1621 gttaaattaa attttactaa agctggttac ttattttttc ccttcaatat aggatttaaa
     1681 ccttgtactt taagaacatg ctataatttg aatagagttt taagatacgt ctttttttct
     1741 atttgggaga ttctaaagta ttaaaatggt cagtaacgtg gttgagatgg tgatttgatg
     1801 attttttttt ttctgtataa gattaaatct aatggtcaag tatttacaat gaattaaggc
     1861 tggttatttt taaaagtctt gctttaagaa tgggaaagtt tttttatcat taaattgaat
     1921 tttgaaatta ttgaaatgct gtaatacaag cataaattag tggtgagatc caagaggcgt
     1981 ttctaatact tttattttct tttcagcgag ggctcagtac ctgaaaacag ccaagcttgg
     2041 caatccggta ctgttggtaa agccaccatg gaagatgcca aaaacattaa gaagggccca
     2101 gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag
     2161 cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt
     2221 acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg
     2281 ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc
     2341 gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag
     2401 cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa
     2461 gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc
     2521 atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat
     2581 ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc
     2641 atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg
     2701 caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc
     2761 atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc
     2821 acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag
     2881 ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta
     2941 tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag
     3001 atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc
     3061 cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc
     3121 acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct
     3181 aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc
     3241 gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc
     3301 atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac
     3361 ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca
     3421 gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc
     3481 ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa
     3541 accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag
     3601 ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac
     3661 gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgtaa
     3721 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     3781 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3841 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3901 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3961 ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa
     4021 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     4081 cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca
     4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4741 gaagaacagt atttggtatc tgcgctctgc
//
