LOCUS       sequence-466694-        5532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near FRMD4A, chr10:14017469-14017970 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5532)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5532)
  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_FRMD4A
FEATURES             Location/Qualifiers
     source          1..5532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(557..1348,1349..1417))
                     /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     1180..1199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1418..1522)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1549..2004
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1636..1655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1846..1867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2135..2183
                     /note="synthetic polyadenylation signal"
     misc_feature    2197..2288
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2237..2256
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2859..3055
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2906..3041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3085..3094
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3091..4743
                     /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     complement(3157..3175)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4784..4905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4795..4814)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4849..4868
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5159..5176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5330..5532,1..386))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5410..5429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
       61 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      121 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      181 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      241 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      301 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      361 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      421 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      481 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      541 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      601 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      661 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      721 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      781 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      841 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
      901 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      961 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1021 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1081 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1141 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1201 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1261 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1321 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1381 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1441 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1501 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     1561 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1621 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1681 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1741 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1801 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1861 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1921 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     1981 acaaaatatt aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag
     2041 ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta
     2101 atattaaggt acgggaggta cttggagcgg ccgcaataaa atatctttat tttcattaca
     2161 tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa
     2221 aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac
     2281 atttctctat cgataggtac cgagctctta cgcgtgctag cgcatacctc caaacagagc
     2341 caatagctgt tgactcaatt tttactaggc ttagctctca aatttcaggt gtcttaataa
     2401 ggattgggtg gcgaatgatc aacttttaca tctgtttcta atttccatgc ttccttttcc
     2461 aggaggagaa atttaacaag caattttctt caacagggga atagttcatc ttggttcaag
     2521 taatccctct ggtgctgcaa aggacaatgg ctttctttag ggccatcaga ttccttttgc
     2581 tcctgtgccc agggacatat cactcccagg gtcaactgtc catttgctca tggacatctg
     2641 ggatgagtca cgtggtatat tttcccaaag tgaggaagga agaaaagaaa gcatcttgaa
     2701 tctgttagaa agacagtctc tttgaagcct catgtcatgg gaatatggtt tatgttgaat
     2761 gtgttctaga gtatacccaa catcctgtaa atcaaaacac caaagtgcaa gctgtgacca
     2821 gaatgggtaa aacacctcca gtctcgagat ctgcgatctg catctcaatt agtcagcaac
     2881 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2941 tccgccccat cgctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3001 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3061 tggcattccg gtactgttgg taaagccacc atggaagacg ccaaaaacat aaagaaaggc
     3121 ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg
     3181 aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac
     3241 atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat
     3301 gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg
     3361 ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat
     3421 gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa
     3481 aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt
     3541 atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct
     3601 catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag
     3661 acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg
     3721 cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa
     3781 atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt
     3841 actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa
     3901 gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc
     3961 ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac
     4021 gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg
     4081 ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc agctattctg
     4141 attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa
     4201 gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg
     4261 tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc
     4321 ttgattgaca aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa
     4381 cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct
     4441 cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca
     4501 ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     4561 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     4621 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     4681 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg
     4741 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     4801 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4861 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4921 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     4981 ctctacaaat gtggtaaaat cgataaggat ctggatccgt cgaccgatgc ccttgagagc
     5041 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     5101 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     5161 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5221 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5281 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5341 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5401 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5461 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5521 ctcatagctc ac
//
