LOCUS       sequence-462845-        5676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near FAM153CP, buffering element replaced with SOX9 site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5676)
  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_FAM153CP_SOX9rep
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    699..790
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(731..750)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    804..852
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(983..1438)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1120..1141)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1332..1351
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1465..1569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1570..1638,1639..2430)
                     /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(1788..1807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2601..3189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3090..3109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3343..3360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3614..3735
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3651..3670)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3705..3724
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3776..5428)
                     /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     5344..5362
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      5425..5434
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5464..5660)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5478..5613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tacaactttt gcctttgtgc tattgctgga atgcaaagct tctacttcag atgagcgctg
       61 agagccaaca aggttgaaat ggaaaaggat ttcctcaatg tctttagctc ccgggtcagg
      121 tagacaagct cggaggtctt caaactgggg gtacacaggt ttctgggtgc acaaagtctt
      181 tctaggaggg aagaaaaata ggttttgcta ctttattgac tagtgaaaag cagcatggtg
      241 gggtgtggct cgcttcttca gtgctgcctc tgctcaaacc tctatgggtg catacagatg
      301 ggcaggctgt ggagctccga ccccacagca ccgtctaggg gtgaatgttc acagctcctg
      361 aagccccagt gggcgtgtgt accatgtgct ctttcagttt agccgcccat acaatgggcc
      421 cattgttcac agctcaatta gacccctgcc ttatcacaag aacaaagggc tttctgtgtc
      481 ctgtggtcct tgctttggtg tagcggaaga attggatcac acatggtctt ggtgaatgag
      541 tgcaaggttt tattgagtag aagtagctct cagatggggg agccagaagg gagatggttt
      601 tctcctggag tcgggccact cggtggcccg agatctcctc caactgcccc agccaaacgc
      661 tacgtgctag cacgcgtaag agctcggtac ctatcgatag agaaatgttc tggcacctgc
      721 acttgcactg gggacagcct attttgctag tttgttttgt ttcgttttgt tttgatggag
      781 agcgtatgtt agtactatcg attcacacaa aaaaccaaca cacagatgta atgaaaataa
      841 agatatttta ttgcggccgc tccaagtacc tcccgtacct taatattact tacttatcat
      901 ggtagcttgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      961 cgcagcctga atggcgaatg gcaaattgta agcgttaata ttttgttaaa attcgcgtta
     1021 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1081 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1141 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1201 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1261 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1321 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1381 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     1441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1801 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2101 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3361 gaagcggaag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     3421 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     3481 tctcaagggc atcggtcgac ggatccagat ccttatcgat tttaccacat ttgtagaggt
     3541 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3601 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3661 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3721 catcaatgta tcttatcatg tctgctcgaa gcggccggcc gccccgactc tagaattaca
     3781 cggcgatctt tccgcccttc ttggccttta tgaggatctc tctgattttt cttgcgtcga
     3841 gttttccggt aagacctttc ggtacttcgt ccacaaacac aactcctccg cgcaactttt
     3901 tcgcggttgt tacttgactg gcgacgtaat ccacgatctc tttttccgtc atcgtctttc
     3961 cgtgctccaa aacaacaacg gcggcgggaa gttcaccggc gtcatcgtcg ggaagacctg
     4021 cgacacctgc gtcgaagatg ttggggtgtt ggagcaagat ggattccaat tcagcgggag
     4081 ccacctgata gcctttgtac ttaatcagag acttcaggcg gtcaacgatg aagaagtgtt
     4141 cgtcttcgtc ccagtaagct atgtctccag aatgtagcca tccatccttg tcaatcaagg
     4201 cgttggtcgc ttccggattg tttacataac cggacataat cataggacct ctcacacaca
     4261 gttcgcctct ttgattaacg cccagcgttt tcccggtatc cagatccaca accttcgctt
     4321 caaaaaatgg aacaacttta ccgaccgcgc ccggtttatc atccccctcg ggtgtaatca
     4381 gaatagctga tgtagtctca gtgagcccat atccttgcct gatacctggc agatggaacc
     4441 tcttggcaac cgcttccccg acttccttag agaggggagc gccaccagaa gcaatttcgt
     4501 gtaaattaga taaatcgtat ttgtcaatca gagtgctttt ggcgaagaag gagaataggg
     4561 ttggcaccag cagcgcactt tgaatcttgt aatcctgaag gctcctcaga aacagctctt
     4621 cttcaaatct atacattaag acgactcgaa atccacatat caaatatccg agtgtagtaa
     4681 acattccaaa accgtgatgg aatggaacaa cacttaaaat cgcagtatcc ggaatgattt
     4741 gattgccaaa aataggatct ctggcatgcg agaatctcac gcaggcagtt ctatgaggca
     4801 gagcgacacc tttaggcaga ccagtagatc cagaggagtt catgatcagt gcaattgtct
     4861 tgtccctatc gaaggactct ggcacaaaat cgtattcatt aaaaccggga ggtagatgag
     4921 atgtgacgaa cgtgtacatc gactgaaatc cctggtaatc cgttttagaa tccatgataa
     4981 taattttttg gatgattggg agcttttttt gcacgttcaa aattttttgc aacccctttt
     5041 tggaaacgaa caccacggta ggctgcgaaa tgcccatact gttgagcaat tcacgttcat
     5101 tataaatgtc gttcgcgggc gcaactgcaa ctccgataaa taacgcgccc aacaccggca
     5161 taaagaattg aagagagttt tcactgcata cgacgattct gtgatttgta ttcagcccat
     5221 atcgtttcat agcttctgcc aaccgaacgg acatttcgaa gtactcagcg taagtgatgt
     5281 ccacctcgat atgtgcatct gtaaaagcaa ttgttccagg aaccagggcg tatctcttca
     5341 tagccttatg cagttgctct ccagcggttc catcttccag cggatagaat ggcgccgggc
     5401 ctttctttat gtttttggcg tcttccatgg tggctttacc aacagtaccg gaatgccaag
     5461 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     5521 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagcgat ggggcggaga
     5581 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5641 tgctgactaa ttgagatgca gatcgcagat ctcgag
//
