LOCUS       sequence-383826-        6699 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 6699)
  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 6699)
  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 E3E2E1-Prg4 promoter-luciferase
FEATURES             Location/Qualifiers
     source          1..6699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(29..617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(109..128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(817..1608,1609..1677))
                     /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    1782..1830
                     /note="synthetic polyadenylation signal"
     misc_feature    1844..1935
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1884..1903
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     RBS             2627..2635
                     /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      4486..4495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4492..6144
                     /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    6188..6309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6199..6218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6253..6272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6557..6574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
       61 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
      661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
      721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
      781 ctgacagcgg ccgcaaatgc taaaccactg cagtggttac cagtgcttga tcagtgaggc
      841 accgatctca gcgatctgcc tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta
      901 gatcactacg attcgtgagg gcttaccatc aggccccagc gcagcaatga tgccgcgaga
      961 gccgcgttca ccggcccccg atttgtcagc aatgaaccag ccagcaggga gggccgagcg
     1021 aagaagtggt cctgctactt tgtccgcctc catccagtct atgagctgct gtcgtgatgc
     1081 tagagtaaga agttcgccag tgagtagttt ccgaagagtt gtggccattg ctactggcat
     1141 cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc agcggtcaag
     1201 ccgggtcaca tgatcaccca tattatgaag aaatgcagtc agctccttag ggcctccgat
     1261 cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa
     1321 ttctcttacc gtcatgccat ccgtaagatg cttttccgtg accggcgagt actcaaccaa
     1381 gtcgttttgt gagtagtgta tacggcgacc aagctgctct tgcccggcgt ctatacggga
     1441 caacaccgcg ccacatagca gtactttgaa agtgctcatc atcgggaatc gttcttcggg
     1501 gcggaaagac tcaaggatct tgccgctatt gagatccagt tcgatatagc ccactcttgc
     1561 acccagttga tcttcagcat cttttacttt caccagcgtt tcggggtgtg caaaaacagg
     1621 caagcaaaat gccgcaaaga agggaatgag tgcgacacga aaatgttgga tgctcatact
     1681 cgtccttttt caatattatt gaagcattta tcagggttac tagtacgtct ctcaaggata
     1741 agtaagtaat attaaggtac gggaggtatt ggacaggccg caataaaata tctttatttt
     1801 cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc
     1861 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     1921 ccagaacatt tctctggcct aactggccgg tacctgagct cgctagcctc gagagggtat
     1981 gtcttgcttt ctcttcccag gcatgctttt gaaaatcctc ctgcctctct ctccttgacc
     2041 aacaagccct gtctgcagtc atgtttctca ctcacaggaa gtcagtaatc cctcaggggc
     2101 taggggttta ctgtgtgttc cttcttcaga gagtgggcaa acaaacgaat tcatcctgca
     2161 gcagcgattg ccagctttga gctgccaggg tttcctggaa caagtctgct cagtgtgtaa
     2221 tgcaccaggc aacatctaga atccctcctc agcagggagc attttatagg gtgatagagc
     2281 cgacctatct ttgaaattgt cttttctgtt ggtgttgcct cagggttttc agggctctct
     2341 gcttttagag cactggctct aagaggtacc acggctgtta cagtgatgag tcatcatgac
     2401 tccatgggag tgtggaggag accaaggatg agtcagtagt ctgtctgttc attcacatct
     2461 catcttgttt acatattctg ctgtctgaaa cactgtcttt tctggttttg aataaactgc
     2521 tttgttctct ccctcccctt acagaaacat tgcactccgt ttaggtacca agttggtggg
     2581 gtgcttggaa aatgagataa tggccatcgt gaataggctc cactaataag gaggtagttc
     2641 tgaaaccgaa ctctggttgg atctcacttt gttttcctct cagtggtgac tcccatgtta
     2701 ctgtaacatg gtcaagtgtg cgctgcccag gattcatcca aaacctattt gtaatctgtc
     2761 tgaggggaat agggagatat cttcaatttt ccattggaga tacgtttctc tctgttttcg
     2821 ctccaaccca gggctcactc cctctgcttc cttctacctt gctttactat gcagaatgcc
     2881 tgaatgctgg ccagcttttc ctttgctaag ggcttatgaa attcttattt caaccaagcg
     2941 gtctctcttg ggaaggccac atgggagtgc aaagtgagtt gtcgcttaaa aaaataaatt
     3001 tttccggatc attgcattac acaatctgta gtttcttgac tttcctgtgt caaactaatt
     3061 ttttagattc aaaagcagag ccacagatgg ttcaactggt ttctcaaaag ttttaaacaa
     3121 tcccaacttt agttttactg gcattcgaat aaattgctct gccttttccc tcctctggta
     3181 ttcctacctg ccttgtactg aataaagata gatggttctg tgtaagactg tgtcaactat
     3241 ttacgtctag atttatcttt tttggctttt tacttctttt ctccttaagt tccccacatt
     3301 ccttttctac aggaagcgtg gaaaagataa aggtgccagg aaagatatta gtttttggct
     3361 gctggcagat ctaatactca ggaaaagaat taaacatttg aatgtattag ttcagaaaac
     3421 ttatgccata agctgaaact agataaagtt tcaaaaacac tctttgctgg gttaactgga
     3481 aaaagaaaaa aaaaaagaaa tgataccagc atttgactat tttaaagttg ttgcactagt
     3541 caaagcctgc caagcaattt accatctcac agaacaattt gttcacttca cttagaaatt
     3601 agaagtgtgt ttgggccaat aaagctaaat ttgtagtagt catttttatt aacggaagtg
     3661 ttttccagat gttacttaag tctggttgtc tgggacctgt tccaataaat gaggaaagtt
     3721 tgatttcata ggttgtcact gttgattctg tctaaccttt ggactaattg gttcatccta
     3781 acatatttgc tccggtatat aagagcattg gacattggag aaattcacct ctccttttac
     3841 gggtaagtgt cacagttaac ttgcttcatg gagttatatg ttttcaattc aataaggatt
     3901 tattaaaagt ttattaaagg tgcttgaaaa gatttataag gatagtagtt ggagagttta
     3961 gaaaagaatc acatttatgt aatacacctg ccggaggctt atgaatttat attgagagtt
     4021 aagagaatgt tggaatgata actgttaaat taaattttac taaagctggt tacttatttt
     4081 ttcccttcaa tataggattt aaaccttgta ctttaagaac atgctataat ttgaatagag
     4141 ttttaagata cgtctttttt tctatttggg agattctaaa gtattaaaat ggtcagtaac
     4201 gtggttgaga tggtgatttg atgatttttt ttttttctgt ataagattaa atctaatggt
     4261 caagtattta caatgaatta aggctggtta tttttaaaag tcttgcttta agaatgggaa
     4321 agttttttta tcattaaatt gaattttgaa attattgaaa tgctgtaata caagcataaa
     4381 ttagtggtga gatccaagag gcgtttctaa tacttttatt ttcttttcag cgagggctca
     4441 gtacctgaaa acagccaagc ttggcaatcc ggtactgttg gtaaagccac catggaagat
     4501 gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg gaccgccggc
     4561 gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat cgcctttacc
     4621 gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag cgttcggctg
     4681 gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag
     4741 aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc
     4801 ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat cagccagccc
     4861 accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca aaagaagcta
     4921 ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg cttccaaagc
     4981 atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga cttcgtgccc
     5041 gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg cagtaccgga
     5101 ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag tcatgcccgc
     5161 gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt ggtgccattt
     5221 caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt tcgggtcgtg
     5281 ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta taagattcaa
     5341 tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct catcgacaag
     5401 tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag caaggaggta
     5461 ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta cggcctgaca
     5521 gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg cgcagtaggc
     5581 aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa gacactgggt
     5641 gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac
     5701 aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag cggcgacatc
     5761 gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag cctgatcaaa
     5821 tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca acaccccaac
     5881 atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct gcccgccgca
     5941 gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga ctatgtggcc
     6001 agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct
     6061 aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat taaggccaag
     6121 aagggcggca agatcgccgt gtaataattc tagagtcggg gcggccggcc gcttcgagca
     6181 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     6241 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     6301 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     6361 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg
     6421 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     6481 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     6541 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6601 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6661 gaaagaacat gtgagcaaaa ggccagcaaa aggccagga
//
