LOCUS       sequence-316810-        6035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is a modified version of the pGL4.23 vector from Promega, 
            containing a Gateway cassette upstream of the minimal promoter. This
            vector can be used for for Gateway cloning of candidate enhancer 
            elements. As negative control, use pGL4.23 without the gateway 
            cassette (available from Promega)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6035)
  AUTHORS   Pasquali L, Gaulton KJ, Rodriguez-Segui SA, Mularoni L, 
            Miguel-Escalada I, Akerman I, Tena JJ, Moran I, Gomez-Marin C, van 
            de Bunt M, Ponsa-Cobas J, Castro N, Nammo T, Cebola I, 
            Garcia-Hurtado J, Maestro MA, Pattou F, Piemonti L, Berney T, Gloyn 
            AL, Ravassard P, Gomez-Skarmeta JL, Muller F, McCarthy MI, Ferrer J
  TITLE     Pancreatic islet enhancer clusters enriched in type 2 diabetes 
            risk-associated variants.
  JOURNAL   Nat Genet. 2014 Feb;46(2):136-43. doi: 10.1038/ng.2870. Epub 2014 
            Jan 12.
  PUBMED    24413736
REFERENCE   2  (bases 1 to 6035)
  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: "10.1038/ng.2870"; 
            journalName: "Nat Genet"; date: "2014-02"; volume: "46"; issue: "2";
            pages: "136-43"
COMMENT     Sequence Label: pGL4.23-GW
FEATURES             Location/Qualifiers
     source          1..6035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(91..96,97..114,115..121)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             175..834
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(241..260)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1176..1481
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1360..1379
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1522..1646)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1720..1751
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1778..1787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1784..3436
                     /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    3480..3601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3491..3510)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3545..3564
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3849..3866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4020..4608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4100..4119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4808..5599,5600..5668))
                     /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    5773..5821
                     /note="synthetic polyadenylation signal"
     misc_feature    5835..5926
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5875..5894
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    join(5977..6035,1..66)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
       61 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
      121 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
      181 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
      241 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
      301 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
      361 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
      421 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
      481 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
      541 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
      601 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
      661 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
      721 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
      781 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
      841 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
      901 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
      961 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     1021 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     1081 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     1141 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     1201 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     1261 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     1321 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     1381 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     1441 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     1501 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     1561 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     1621 cgttcagctt tcttgtacaa agtggtgata aacccgctga tcagcctcga ctgtgctcga
     1681 ggatatcaag atctggcctc ggcggccaag cttagacact agagggtata taatggaagc
     1741 tcgacttcca gcttggcaat ccggtactgt tggtaaagcc accatggaag atgccaaaaa
     1801 cattaagaag ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct
     1861 gcacaaagcc atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca
     1921 tatcgaggtg gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc
     1981 tatgaagcgc tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt
     2041 gcagttcttc atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa
     2101 cgacatctac aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt
     2161 attcgtgagc aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat
     2221 acaaaagatc atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac
     2281 cttcgtgact tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt
     2341 cgaccgggac aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa
     2401 gggcgtagcc ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat
     2461 cttcggcaac cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg
     2521 cttcggcatg ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta
     2581 ccgcttcgag gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct
     2641 gctggtgccc acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct
     2701 aagcaacttg cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc
     2761 cgtggccaaa cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac
     2821 cagcgccatt ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt
     2881 gcccttcttc gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca
     2941 gcgcggcgag ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga
     3001 ggctacaaac gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg
     3061 ggacgaggac gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg
     3121 ctaccaggta gccccagccg aactggagag catcctgctg caacacccca acatcttcga
     3181 cgccggggtc gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt
     3241 gctggaacac ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     3301 tacaaccgcc aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact
     3361 gaccggcaag ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg
     3421 caagatcgcc gtgtaataat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     3481 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3541 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3601 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3661 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     3721 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     3781 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     3841 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3901 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3961 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4021 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4081 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4141 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4201 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4261 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4321 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4381 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4441 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4501 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4561 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4621 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4681 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4741 tcaatctaaa gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact
     4801 gcagtggtta ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt
     4861 cgtccatagt ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat
     4921 caggccccag cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag
     4981 caatgaacca gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct
     5041 ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt
     5101 tccgaagagt tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg
     5161 cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa
     5221 gaaatgcagt cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt
     5281 tgtcgctcat ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat
     5341 gcttttccgt gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac
     5401 caagctgctc ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga
     5461 aagtgctcat catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat
     5521 tgagatccag ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt
     5581 tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga
     5641 gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt
     5701 atcagggtta ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat
     5761 tggacaggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     5821 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     5881 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctggcc taactggccg
     5941 gtacctgagc tcccgtcgac gaattctgca gatatcacaa gtttgtacaa aaaagctgaa
     6001 cgagaaacgt aaaatgatat aaatatcaat atatt
//
