LOCUS       sequence-462855-        5594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near ST3GAL4, all buffering elements replaced with SOX9 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5594)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5594)
  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"
COMMENT     Sequence Label: pGL3_ST3GAL4_SOX9repall
FEATURES             Location/Qualifiers
     source          1..5594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    617..708
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(649..668)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    722..770
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(901..1356)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1038..1059)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1250..1269
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1383..1487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1488..1556,1557..2348)
                     /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(1706..1725)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2519..3107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3008..3027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3261..3278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3532..3653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3569..3588)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3623..3642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3694..5346)
                     /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     5262..5280
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      5343..5352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5382..5578)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5396..5531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gaccagcact tccccctcct gctcccaccc tctcctcctc tctggccctc gtctcctgcc
       61 ctctcctccc ggtccctgtg tcctttccag ccatcccatc ctggtgtctc ctggtcccca
      121 tctccacctc ctcctccttt cctcctctgc ccacagcccc tggttatgta aggggcttgg
      181 gaacacacag agggagcttt gttgtcaaca atgggcccat tgtagccgga atgggccagc
      241 gacaatgggg ccattgtcct ctgccttggc agggttggga ctccgggcag ggagcccctg
      301 caccccgcca ttttgggaga tgggggacaa acaacaatgg gaccattgtc aggcagctga
      361 atgagtggtc agggatcagt ctccactgtg tggaaccaca gcaggggaca caatgggtcc
      421 attgtcagaa caaagccaca gtacaaagcg agggtcacaa agccaacaga ggcaagctct
      481 gggctggggc agggcgacac ggtgccgggc ctgggagggt ggggagtgta tccagaccct
      541 cacacagaac actgttgtgt gctcacctca atggggtctg agagctagca cgcgtaagag
      601 ctcggtacct atcgatagag aaatgttctg gcacctgcac ttgcactggg gacagcctat
      661 tttgctagtt tgttttgttt cgttttgttt tgatggagag cgtatgttag tactatcgat
      721 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccgctc
      781 caagtacctc ccgtacctta atattactta cttatcatgg tagcttgggc tggcgtaata
      841 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      901 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
      961 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     1021 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     1081 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     1141 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     1201 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     1261 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     1321 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     1381 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1441 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1501 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1561 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1621 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1681 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1741 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1801 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1861 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1921 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1981 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2041 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2101 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2161 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2221 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2281 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2341 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2401 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2461 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2521 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2581 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2641 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2701 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2761 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2821 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2881 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2941 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3001 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3061 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3121 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3181 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3241 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgctgccggc
     3301 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     3361 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgacgg
     3421 atccagatcc ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3481 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3541 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3601 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3661 tgctcgaagc ggccggccgc cccgactcta gaattacacg gcgatctttc cgcccttctt
     3721 ggcctttatg aggatctctc tgatttttct tgcgtcgagt tttccggtaa gacctttcgg
     3781 tacttcgtcc acaaacacaa ctcctccgcg caactttttc gcggttgtta cttgactggc
     3841 gacgtaatcc acgatctctt tttccgtcat cgtctttccg tgctccaaaa caacaacggc
     3901 ggcgggaagt tcaccggcgt catcgtcggg aagacctgcg acacctgcgt cgaagatgtt
     3961 ggggtgttgg agcaagatgg attccaattc agcgggagcc acctgatagc ctttgtactt
     4021 aatcagagac ttcaggcggt caacgatgaa gaagtgttcg tcttcgtccc agtaagctat
     4081 gtctccagaa tgtagccatc catccttgtc aatcaaggcg ttggtcgctt ccggattgtt
     4141 tacataaccg gacataatca taggacctct cacacacagt tcgcctcttt gattaacgcc
     4201 cagcgttttc ccggtatcca gatccacaac cttcgcttca aaaaatggaa caactttacc
     4261 gaccgcgccc ggtttatcat ccccctcggg tgtaatcaga atagctgatg tagtctcagt
     4321 gagcccatat ccttgcctga tacctggcag atggaacctc ttggcaaccg cttccccgac
     4381 ttccttagag aggggagcgc caccagaagc aatttcgtgt aaattagata aatcgtattt
     4441 gtcaatcaga gtgcttttgg cgaagaagga gaatagggtt ggcaccagca gcgcactttg
     4501 aatcttgtaa tcctgaaggc tcctcagaaa cagctcttct tcaaatctat acattaagac
     4561 gactcgaaat ccacatatca aatatccgag tgtagtaaac attccaaaac cgtgatggaa
     4621 tggaacaaca cttaaaatcg cagtatccgg aatgatttga ttgccaaaaa taggatctct
     4681 ggcatgcgag aatctcacgc aggcagttct atgaggcaga gcgacacctt taggcagacc
     4741 agtagatcca gaggagttca tgatcagtgc aattgtcttg tccctatcga aggactctgg
     4801 cacaaaatcg tattcattaa aaccgggagg tagatgagat gtgacgaacg tgtacatcga
     4861 ctgaaatccc tggtaatccg ttttagaatc catgataata attttttgga tgattgggag
     4921 ctttttttgc acgttcaaaa ttttttgcaa cccctttttg gaaacgaaca ccacggtagg
     4981 ctgcgaaatg cccatactgt tgagcaattc acgttcatta taaatgtcgt tcgcgggcgc
     5041 aactgcaact ccgataaata acgcgcccaa caccggcata aagaattgaa gagagttttc
     5101 actgcatacg acgattctgt gatttgtatt cagcccatat cgtttcatag cttctgccaa
     5161 ccgaacggac atttcgaagt actcagcgta agtgatgtcc acctcgatat gtgcatctgt
     5221 aaaagcaatt gttccaggaa ccagggcgta tctcttcata gccttatgca gttgctctcc
     5281 agcggttcca tcttccagcg gatagaatgg cgccgggcct ttctttatgt ttttggcgtc
     5341 ttccatggtg gctttaccaa cagtaccgga atgccaagct ttttgcaaaa gcctaggcct
     5401 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     5461 gcataaataa aaaaaattag tcagcgatgg ggcggagaat gggcggaact gggcggagtt
     5521 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcaga
     5581 tcgcagatct cgag
//
