LOCUS       sequence-383827-        7030 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 7030)
  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 7030)
  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: pGL4.10 E4E3E2E1-Prg4 promoter-luciferase
FEATURES             Location/Qualifiers
     source          1..7030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(357..1148,1149..1217))
                     /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    1322..1370
                     /note="synthetic polyadenylation signal"
     misc_feature    1384..1475
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1424..1443
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     RBS             2498..2506
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      4357..4366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4363..6015
                     /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    6059..6180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6070..6089)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6124..6143
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6428..6445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6599..7030,1..157))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6679..6698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
       61 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      301 tatatatgag taaacttggt ctgacagcgg ccgcaaatgc taaaccactg cagtggttac
      361 cagtgcttga tcagtgaggc accgatctca gcgatctgcc tatttcgttc gtccatagtg
      421 gcctgactcc ccgtcgtgta gatcactacg attcgtgagg gcttaccatc aggccccagc
      481 gcagcaatga tgccgcgaga gccgcgttca ccggcccccg atttgtcagc aatgaaccag
      541 ccagcaggga gggccgagcg aagaagtggt cctgctactt tgtccgcctc catccagtct
      601 atgagctgct gtcgtgatgc tagagtaaga agttcgccag tgagtagttt ccgaagagtt
      661 gtggccattg ctactggcat cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac
      721 tctggttccc agcggtcaag ccgggtcaca tgatcaccca tattatgaag aaatgcagtc
      781 agctccttag ggcctccgat cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg
      841 gtaatggcag cactacacaa ttctcttacc gtcatgccat ccgtaagatg cttttccgtg
      901 accggcgagt actcaaccaa gtcgttttgt gagtagtgta tacggcgacc aagctgctct
      961 tgcccggcgt ctatacggga caacaccgcg ccacatagca gtactttgaa agtgctcatc
     1021 atcgggaatc gttcttcggg gcggaaagac tcaaggatct tgccgctatt gagatccagt
     1081 tcgatatagc ccactcttgc acccagttga tcttcagcat cttttacttt caccagcgtt
     1141 tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga agggaatgag tgcgacacga
     1201 aaatgttgga tgctcatact cgtccttttt caatattatt gaagcattta tcagggttac
     1261 tagtacgtct ctcaaggata agtaagtaat attaaggtac gggaggtatt ggacaggccg
     1321 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     1381 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     1441 ccccagtgca agtgcaggtg ccagaacatt tctctggcct aactggccgg tacctgagct
     1501 cgctagcgga aaactattaa tcagcatagt ggaggaacat ttcttaactc ttcaatgttt
     1561 tcattggaaa gcattccaaa aataacggag acaacaccct tgtaattaaa atttccaagt
     1621 gcagagttcc tgcagctttc agatggacct atgagatgac cttcttattc actcatcatt
     1681 ctgataattg ttttggctgt gagacaaaaa actaagaatt tattttgggc agaggatgga
     1741 tcaggaggga agatgattaa agaatgatca atcattgaga gaaccatatt aataaaggca
     1801 atagtttatg ccaccagggg cgggtcagat tacttgagct cgagagggta tgtcttgctt
     1861 tctcttccca ggcatgcttt tgaaaatcct cctgcctctc tctccttgac caacaagccc
     1921 tgtctgcagt catgtttctc actcacagga agtcagtaat ccctcagggg ctaggggttt
     1981 actgtgtgtt ccttcttcag agagtgggca aacaaacgaa ttcatcctgc agcagcgatt
     2041 gccagctttg agctgccagg gtttcctgga acaagtctgc tcagtgtgta atgcaccagg
     2101 caacatctag aatccctcct cagcagggag cattttatag ggtgatagag ccgacctatc
     2161 tttgaaattg tcttttctgt tggtgttgcc tcagggtttt cagggctctc tgcttttaga
     2221 gcactggctc taagaggtac cacggctgtt acagtgatga gtcatcatga ctccatggga
     2281 gtgtggagga gaccaaggat gagtcagtag tctgtctgtt cattcacatc tcatcttgtt
     2341 tacatattct gctgtctgaa acactgtctt ttctggtttt gaataaactg ctttgttctc
     2401 tccctcccct tacagaaaca ttgcactccg tttaggtacc aagttggtgg ggtgcttgga
     2461 aaatgagata atggccatcg tgaataggct ccactaataa ggaggtagtt ctgaaaccga
     2521 actctggttg gatctcactt tgttttcctc tcagtggtga ctcccatgtt actgtaacat
     2581 ggtcaagtgt gcgctgccca ggattcatcc aaaacctatt tgtaatctgt ctgaggggaa
     2641 tagggagata tcttcaattt tccattggag atacgtttct ctctgttttc gctccaaccc
     2701 agggctcact ccctctgctt ccttctacct tgctttacta tgcagaatgc ctgaatgctg
     2761 gccagctttt cctttgctaa gggcttatga aattcttatt tcaaccaagc ggtctctctt
     2821 gggaaggcca catgggagtg caaagtgagt tgtcgcttaa aaaaataaat ttttccggat
     2881 cattgcatta cacaatctgt agtttcttga ctttcctgtg tcaaactaat tttttagatt
     2941 caaaagcaga gccacagatg gttcaactgg tttctcaaaa gttttaaaca atcccaactt
     3001 tagttttact ggcattcgaa taaattgctc tgccttttcc ctcctctggt attcctacct
     3061 gccttgtact gaataaagat agatggttct gtgtaagact gtgtcaacta tttacgtcta
     3121 gatttatctt ttttggcttt ttacttcttt tctccttaag ttccccacat tccttttcta
     3181 caggaagcgt ggaaaagata aaggtgccag gaaagatatt agtttttggc tgctggcaga
     3241 tctaatactc aggaaaagaa ttaaacattt gaatgtatta gttcagaaaa cttatgccat
     3301 aagctgaaac tagataaagt ttcaaaaaca ctctttgctg ggttaactgg aaaaagaaaa
     3361 aaaaaaagaa atgataccag catttgacta ttttaaagtt gttgcactag tcaaagcctg
     3421 ccaagcaatt taccatctca cagaacaatt tgttcacttc acttagaaat tagaagtgtg
     3481 tttgggccaa taaagctaaa tttgtagtag tcatttttat taacggaagt gttttccaga
     3541 tgttacttaa gtctggttgt ctgggacctg ttccaataaa tgaggaaagt ttgatttcat
     3601 aggttgtcac tgttgattct gtctaacctt tggactaatt ggttcatcct aacatatttg
     3661 ctccggtata taagagcatt ggacattgga gaaattcacc tctcctttta cgggtaagtg
     3721 tcacagttaa cttgcttcat ggagttatat gttttcaatt caataaggat ttattaaaag
     3781 tttattaaag gtgcttgaaa agatttataa ggatagtagt tggagagttt agaaaagaat
     3841 cacatttatg taatacacct gccggaggct tatgaattta tattgagagt taagagaatg
     3901 ttggaatgat aactgttaaa ttaaatttta ctaaagctgg ttacttattt tttcccttca
     3961 atataggatt taaaccttgt actttaagaa catgctataa tttgaataga gttttaagat
     4021 acgtcttttt ttctatttgg gagattctaa agtattaaaa tggtcagtaa cgtggttgag
     4081 atggtgattt gatgattttt tttttttctg tataagatta aatctaatgg tcaagtattt
     4141 acaatgaatt aaggctggtt atttttaaaa gtcttgcttt aagaatggga aagttttttt
     4201 atcattaaat tgaattttga aattattgaa atgctgtaat acaagcataa attagtggtg
     4261 agatccaaga ggcgtttcta atacttttat tttcttttca gcgagggctc agtacctgaa
     4321 aacagccaag cttggcaatc cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac
     4381 attaagaagg gcccagcgcc attctaccca ctcgaagacg ggaccgccgg cgagcagctg
     4441 cacaaagcca tgaagcgcta cgccctggtg cccggcacca tcgcctttac cgacgcacat
     4501 atcgaggtgg acattaccta cgccgagtac ttcgagatga gcgttcggct ggcagaagct
     4561 atgaagcgct atgggctgaa tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg
     4621 cagttcttca tgcccgtgtt gggtgccctg ttcatcggtg tggctgtggc cccagctaac
     4681 gacatctaca acgagcgcga gctgctgaac agcatgggca tcagccagcc caccgtcgta
     4741 ttcgtgagca agaaagggct gcaaaagatc ctcaacgtgc aaaagaagct accgatcata
     4801 caaaagatca tcatcatgga tagcaagacc gactaccagg gcttccaaag catgtacacc
     4861 ttcgtgactt cccatttgcc acccggcttc aacgagtacg acttcgtgcc cgagagcttc
     4921 gaccgggaca aaaccatcgc cctgatcatg aacagtagtg gcagtaccgg attgcccaag
     4981 ggcgtagccc taccgcaccg caccgcttgt gtccgattca gtcatgcccg cgaccccatc
     5041 ttcggcaacc agatcatccc cgacaccgct atcctcagcg tggtgccatt tcaccacggc
     5101 ttcggcatgt tcaccacgct gggctacttg atctgcggct ttcgggtcgt gctcatgtac
     5161 cgcttcgagg aggagctatt cttgcgcagc ttgcaagact ataagattca atctgccctg
     5221 ctggtgccca cactatttag cttcttcgct aagagcactc tcatcgacaa gtacgaccta
     5281 agcaacttgc acgagatcgc cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc
     5341 gtggccaaac gcttccacct accaggcatc cgccagggct acggcctgac agaaacaacc
     5401 agcgccattc tgatcacccc cgaaggggac gacaagcctg gcgcagtagg caaggtggtg
     5461 cccttcttcg aggctaaggt ggtggacttg gacaccggta agacactggg tgtgaaccag
     5521 cgcggcgagc tgtgcgtccg tggccccatg atcatgagcg gctacgttaa caaccccgag
     5581 gctacaaacg ctctcatcga caaggacggc tggctgcaca gcggcgacat cgcctactgg
     5641 gacgaggacg agcacttctt catcgtggac cggctgaaga gcctgatcaa atacaagggc
     5701 taccaggtag ccccagccga actggagagc atcctgctgc aacaccccaa catcttcgac
     5761 gccggggtcg ccggcctgcc cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg
     5821 ctggaacacg gtaaaaccat gaccgagaag gagatcgtgg actatgtggc cagccaggtt
     5881 acaaccgcca agaagctgcg cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg
     5941 accggcaagt tggacgcccg caagatccgc gagattctca ttaaggccaa gaagggcggc
     6001 aagatcgccg tgtaataatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     6061 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     6121 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     6181 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     6241 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc
     6301 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     6361 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     6421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6781 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7021 actacggcta
//
