LOCUS       sequence-383820-        5897 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 5897)
  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 5897)
  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 E2E1- Prg4 promoter-luciferase
FEATURES             Location/Qualifiers
     source          1..5897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(77..868,869..937))
                     /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    1042..1090
                     /note="synthetic polyadenylation signal"
     misc_feature    1104..1195
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1144..1163
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2944..2953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2950..4602
                     /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    4646..4767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4657..4676)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4711..4730
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5015..5032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5186..5774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5266..5285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc
       61 taaaccactg cagtggttac cagtgcttga tcagtgaggc accgatctca gcgatctgcc
      121 tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta gatcactacg attcgtgagg
      181 gcttaccatc aggccccagc gcagcaatga tgccgcgaga gccgcgttca ccggcccccg
      241 atttgtcagc aatgaaccag ccagcaggga gggccgagcg aagaagtggt cctgctactt
      301 tgtccgcctc catccagtct atgagctgct gtcgtgatgc tagagtaaga agttcgccag
      361 tgagtagttt ccgaagagtt gtggccattg ctactggcat cgtggtatca cgctcgtcgt
      421 tcggtatggc ttcgttcaac tctggttccc agcggtcaag ccgggtcaca tgatcaccca
      481 tattatgaag aaatgcagtc agctccttag ggcctccgat cgttgtcaga agtaagttgg
      541 ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa ttctcttacc gtcatgccat
      601 ccgtaagatg cttttccgtg accggcgagt actcaaccaa gtcgttttgt gagtagtgta
      661 tacggcgacc aagctgctct tgcccggcgt ctatacggga caacaccgcg ccacatagca
      721 gtactttgaa agtgctcatc atcgggaatc gttcttcggg gcggaaagac tcaaggatct
      781 tgccgctatt gagatccagt tcgatatagc ccactcttgc acccagttga tcttcagcat
      841 cttttacttt caccagcgtt tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga
      901 agggaatgag tgcgacacga aaatgttgga tgctcatact cgtccttttt caatattatt
      961 gaagcattta tcagggttac tagtacgtct ctcaaggata agtaagtaat attaaggtac
     1021 gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct gtgtgttggt
     1081 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     1141 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctggcct
     1201 aactggccgg tacctgagct cgctagcctc gaggatatct tcaattttcc attggagata
     1261 cgtttctctc tgttttcgct ccaacccagg gctcactccc tctgcttcct tctaccttgc
     1321 tttactatgc agaatgcctg aatgctggcc agcttttcct ttgctaaggg cttatgaaat
     1381 tcttatttca accaagcggt ctctcttggg aaggccacat gggagtgcaa agtgagttgt
     1441 cgcttaaaaa aataaatttt tccggatcat tgcattacac aatctgtagt ttcttgactt
     1501 tcctgtgtca aactaatttt ttagattcaa aagcagagcc acagatggtt caactggttt
     1561 ctcaaaagtt ttaaacaatc ccaactttag ttttactggc attcgaataa attgctctgc
     1621 cttttccctc ctctggtatt cctacctgcc ttgtactgaa taaagataga tggttctgtg
     1681 taagactgtg tcaactattt acgtctagat ttatcttttt tggcttttta cttcttttct
     1741 ccttaagttc cccacattcc ttttctacag gaagcgtgga aaagataaag gtgccaggaa
     1801 agatattagt ttttggctgc tggcagatct aatactcagg aaaagaatta aacatttgaa
     1861 tgtattagtt cagaaaactt atgccataag ctgaaactag ataaagtttc aaaaacactc
     1921 tttgctgggt taactggaaa aagaaaaaaa aaaagaaatg ataccagcat ttgactattt
     1981 taaagttgtt gcactagtca aagcctgcca agcaatttac catctcacag aacaatttgt
     2041 tcacttcact tagaaattag aagtgtgttt gggccaataa agctaaattt gtagtagtca
     2101 tttttattaa cggaagtgtt ttccagatgt tacttaagtc tggttgtctg ggacctgttc
     2161 caataaatga ggaaagtttg atttcatagg ttgtcactgt tgattctgtc taacctttgg
     2221 actaattggt tcatcctaac atatttgctc cggtatataa gagcattgga cattggagaa
     2281 attcacctct ccttttacgg gtaagtgtca cagttaactt gcttcatgga gttatatgtt
     2341 ttcaattcaa taaggattta ttaaaagttt attaaaggtg cttgaaaaga tttataagga
     2401 tagtagttgg agagtttaga aaagaatcac atttatgtaa tacacctgcc ggaggcttat
     2461 gaatttatat tgagagttaa gagaatgttg gaatgataac tgttaaatta aattttacta
     2521 aagctggtta cttatttttt cccttcaata taggatttaa accttgtact ttaagaacat
     2581 gctataattt gaatagagtt ttaagatacg tctttttttc tatttgggag attctaaagt
     2641 attaaaatgg tcagtaacgt ggttgagatg gtgatttgat gatttttttt ttttctgtat
     2701 aagattaaat ctaatggtca agtatttaca atgaattaag gctggttatt tttaaaagtc
     2761 ttgctttaag aatgggaaag tttttttatc attaaattga attttgaaat tattgaaatg
     2821 ctgtaataca agcataaatt agtggtgaga tccaagaggc gtttctaata cttttatttt
     2881 cttttcagcg agggctcagt acctgaaaac agccaagctt ggcaatccgg tactgttggt
     2941 aaagccacca tggaagatgc caaaaacatt aagaagggcc cagcgccatt ctacccactc
     3001 gaagacggga ccgccggcga gcagctgcac aaagccatga agcgctacgc cctggtgccc
     3061 ggcaccatcg cctttaccga cgcacatatc gaggtggaca ttacctacgc cgagtacttc
     3121 gagatgagcg ttcggctggc agaagctatg aagcgctatg ggctgaatac aaaccatcgg
     3181 atcgtggtgt gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg tgccctgttc
     3241 atcggtgtgg ctgtggcccc agctaacgac atctacaacg agcgcgagct gctgaacagc
     3301 atgggcatca gccagcccac cgtcgtattc gtgagcaaga aagggctgca aaagatcctc
     3361 aacgtgcaaa agaagctacc gatcatacaa aagatcatca tcatggatag caagaccgac
     3421 taccagggct tccaaagcat gtacaccttc gtgacttccc atttgccacc cggcttcaac
     3481 gagtacgact tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct gatcatgaac
     3541 agtagtggca gtaccggatt gcccaagggc gtagccctac cgcaccgcac cgcttgtgtc
     3601 cgattcagtc atgcccgcga ccccatcttc ggcaaccaga tcatccccga caccgctatc
     3661 ctcagcgtgg tgccatttca ccacggcttc ggcatgttca ccacgctggg ctacttgatc
     3721 tgcggctttc gggtcgtgct catgtaccgc ttcgaggagg agctattctt gcgcagcttg
     3781 caagactata agattcaatc tgccctgctg gtgcccacac tatttagctt cttcgctaag
     3841 agcactctca tcgacaagta cgacctaagc aacttgcacg agatcgccag cggcggggcg
     3901 ccgctcagca aggaggtagg tgaggccgtg gccaaacgct tccacctacc aggcatccgc
     3961 cagggctacg gcctgacaga aacaaccagc gccattctga tcacccccga aggggacgac
     4021 aagcctggcg cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt ggacttggac
     4081 accggtaaga cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg ccccatgatc
     4141 atgagcggct acgttaacaa ccccgaggct acaaacgctc tcatcgacaa ggacggctgg
     4201 ctgcacagcg gcgacatcgc ctactgggac gaggacgagc acttcttcat cgtggaccgg
     4261 ctgaagagcc tgatcaaata caagggctac caggtagccc cagccgaact ggagagcatc
     4321 ctgctgcaac accccaacat cttcgacgcc ggggtcgccg gcctgcccga cgacgatgcc
     4381 ggcgagctgc ccgccgcagt cgtcgtgctg gaacacggta aaaccatgac cgagaaggag
     4441 atcgtggact atgtggccag ccaggttaca accgccaaga agctgcgcgg tggtgttgtg
     4501 ttcgtggacg aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa gatccgcgag
     4561 attctcatta aggccaagaa gggcggcaag atcgccgtgt aataattcta gagtcggggc
     4621 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     4681 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4741 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4801 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     4861 atcgataagg atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     4921 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     4981 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5041 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5101 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5761 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5881 ttttaaatta aaaatga
//
