LOCUS       sequence-425844-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase enhancer reporter plasmid with minimal enhancer 
            regions 1 and 2 (min1 and min2) from SOX9 enhancer cluster EC1.45 
            with E-boxes mutated.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  AUTHORS   Kim S, Morgunova E, Naqvi S, Goovaerts S, Bader M, Koska M, Popov A,
            Luong C, Pogson A, Swigut T, Claes P, Taipale J, Wysocka J
  TITLE     DNA-guided transcription factor cooperativity shapes face and limb 
            mesenchyme.
  JOURNAL   Cell. 2024 Feb 1;187(3):692-711.e26. doi: 
            10.1016/j.cell.2023.12.032. Epub 2024 Jan 22.
  PUBMED    38262408
REFERENCE   2  (bases 1 to 5507)
  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"; doi: 
            "10.1016/j.cell.2023.12.032"; journalName: "Cell"; date: "2024-02-1-
            1"; volume: "187"; issue: "3"; pages: "692-711.e26"
COMMENT     Sequence Label: pGL3-noSV40-humanEC1.45_min1-2_4XEboxMutant
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(32..228)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      46..181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    774..865
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(806..825)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    879..927
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(1058..1513)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1195..1216)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1407..1426
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1540..1644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1645..1713,1714..2505)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1863..1882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2676..3264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3165..3184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3418..3435
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3689..3810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3726..3745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3780..3799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3851..5503)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     5419..5437
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      join(5500..5507,1..2)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gctttaccaa cagtaccgga atgccaagct ttttgcaaaa gcctaggcct ccaaaaaagc
       61 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
      121 aaaaaattag tcagcgatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
      181 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcaga tcgcagatct
      241 ttcactgagt aaggaggaat gaattcattt gtaccattta aatgtgagga aaaggattgc
      301 aaaatcttgg tggtctgggt gggctcaatg tgctcaatat ccattaagat ccaaaattta
      361 ctcgaaccta cagattaatt ctcctgcagg aactgaggca aaccaatatt gagcaaaagt
      421 taatcaagca ctgatctctc ccaatccttc taatcacagt tgagcatgat ttcaaacctt
      481 tgcactgagg ctcgagctgt ttaaaaatag tgaatccctg gcagtagtaa gaataaaggg
      541 tactgttagt gattcaaaaa tgggaaaatt ttctgaaagc acagatgctc aatctaagtt
      601 agatagataa actagaacag aagtgaaacc tgaggctggg tcgatgctcc tttcttccac
      661 actctagagg ggctggttgc aaggttcaac ggggttagcg cggaggcgac ccggctatga
      721 ggcaggaagc atccgcgtcc acggaagagc agctacatgg cgtacctatc gatagagaaa
      781 tgttctggca cctgcacttg cactggggac agcctatttt gctagtttgt tttgtttcgt
      841 tttgttttga tggagagcgt atgttagtac tatcgattca cacaaaaaac caacacacag
      901 atgtaatgaa aataaagata ttttattgcg gccgctccaa gtacctcccg taccttaata
      961 ttacttactt atcatggtag cttgggctgg cgtaatagcg aagaggcccg caccgatcgc
     1021 ccttcccaac agttgcgcag cctgaatggc gaatggcaaa ttgtaagcgt taatattttg
     1081 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1141 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1201 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1261 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1321 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     1381 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     1441 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     1501 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1561 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1621 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1681 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1741 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1801 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1861 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1921 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1981 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2041 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2101 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2161 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2221 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2281 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2341 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2401 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2461 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2521 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2581 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2641 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2701 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2761 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2821 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2881 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2941 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3001 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3061 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3121 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3181 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3241 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3301 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3361 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3421 gagtcagtga gcgaggaagc ggaagagcgc tgccggcacc tgtcctacga gttgcatgat
     3481 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     3541 gactgggttg aaggctctca agggcatcgg tcgacggatc cagatcctta tcgattttac
     3601 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3661 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3721 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3781 tggtttgtcc aaactcatca atgtatctta tcatgtctgc tcgaagcggc cggccgcccc
     3841 gactctagaa ttacacggcg atctttccgc ccttcttggc ctttatgagg atctctctga
     3901 tttttcttgc gtcgagtttt ccggtaagac ctttcggtac ttcgtccaca aacacaactc
     3961 ctccgcgcaa ctttttcgcg gttgttactt gactggcgac gtaatccacg atctcttttt
     4021 ccgtcatcgt ctttccgtgc tccaaaacaa caacggcggc gggaagttca ccggcgtcat
     4081 cgtcgggaag acctgcgaca cctgcgtcga agatgttggg gtgttggagc aagatggatt
     4141 ccaattcagc gggagccacc tgatagcctt tgtacttaat cagagacttc aggcggtcaa
     4201 cgatgaagaa gtgttcgtct tcgtcccagt aagctatgtc tccagaatgt agccatccat
     4261 ccttgtcaat caaggcgttg gtcgcttccg gattgtttac ataaccggac ataatcatag
     4321 gacctctcac acacagttcg cctctttgat taacgcccag cgttttcccg gtatccagat
     4381 ccacaacctt cgcttcaaaa aatggaacaa ctttaccgac cgcgcccggt ttatcatccc
     4441 cctcgggtgt aatcagaata gctgatgtag tctcagtgag cccatatcct tgcctgatac
     4501 ctggcagatg gaacctcttg gcaaccgctt ccccgacttc cttagagagg ggagcgccac
     4561 cagaagcaat ttcgtgtaaa ttagataaat cgtatttgtc aatcagagtg cttttggcga
     4621 agaaggagaa tagggttggc accagcagcg cactttgaat cttgtaatcc tgaaggctcc
     4681 tcagaaacag ctcttcttca aatctataca ttaagacgac tcgaaatcca catatcaaat
     4741 atccgagtgt agtaaacatt ccaaaaccgt gatggaatgg aacaacactt aaaatcgcag
     4801 tatccggaat gatttgattg ccaaaaatag gatctctggc atgcgagaat ctcacgcagg
     4861 cagttctatg aggcagagcg acacctttag gcagaccagt agatccagag gagttcatga
     4921 tcagtgcaat tgtcttgtcc ctatcgaagg actctggcac aaaatcgtat tcattaaaac
     4981 cgggaggtag atgagatgtg acgaacgtgt acatcgactg aaatccctgg taatccgttt
     5041 tagaatccat gataataatt ttttggatga ttgggagctt tttttgcacg ttcaaaattt
     5101 tttgcaaccc ctttttggaa acgaacacca cggtaggctg cgaaatgccc atactgttga
     5161 gcaattcacg ttcattataa atgtcgttcg cgggcgcaac tgcaactccg ataaataacg
     5221 cgcccaacac cggcataaag aattgaagag agttttcact gcatacgacg attctgtgat
     5281 ttgtattcag cccatatcgt ttcatagctt ctgccaaccg aacggacatt tcgaagtact
     5341 cagcgtaagt gatgtccacc tcgatatgtg catctgtaaa agcaattgtt ccaggaacca
     5401 gggcgtatct cttcatagcc ttatgcagtt gctctccagc ggttccatct tccagcggat
     5461 agaatggcgc cgggcctttc tttatgtttt tggcgtcttc catggtg
//
