LOCUS       sequence-466714-        5513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near INPP5F, chr10:119693159-119693660 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5513)
  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_INPP5F
FEATURES             Location/Qualifiers
     source          1..5513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(543..1334,1335..1403))
                     /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     1166..1185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1404..1508)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1535..1990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1622..1641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1832..1853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2121..2169
                     /note="synthetic polyadenylation signal"
     misc_feature    2183..2274
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2223..2242
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             complement(2783..2794)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        2826..3022
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2873..3008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3052..3061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3058..4710
                     /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(3124..3142)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4751..4872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4762..4781)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4816..4835
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5126..5143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5297..5513,1..372))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5377..5396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
       61 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      121 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      181 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      241 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      301 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      361 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
      421 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
      481 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
      541 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
      601 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
      661 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
      721 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
      781 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
      841 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
      901 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
      961 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1021 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1081 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1141 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1201 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1261 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1321 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1381 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     1441 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1501 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     1561 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1621 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1681 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1741 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1801 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1861 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1921 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1981 cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     2041 gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg
     2101 gaggtacttg gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     2161 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     2221 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat
     2281 aggtaccgag ctcttacgcg tgctagctct ctgaccatct ctgccatttc caccactctc
     2341 tacttaccca tcctacacca tcccataatg accactccag cttcagtcaa cttcaggatc
     2401 tttagcccag ctcccaccgt caacagacag atgctcaaga gatggaaccc aactagcctg
     2461 gtaatttctt caagtgagga gacaagtcat aggtcactgg cctcagatta caggatgtgg
     2521 ctgtttagga ccacaggatg gtggacagga cggcctccct gagaggcggt ggggagagca
     2581 gaccacaaga agcagctttg tatgcctccc attcacgcct ccacccaagg catggcctgt
     2641 gtgctcagca gcccggaaac tgctttcact gaggtcataa acagtgtctt tgttgttgaa
     2701 tccaatggac agtattcctt tctttctgtg aatgaatgtt cagtggcttc tggagctgtt
     2761 gacccttttc ctgacccctt tcctgtaact cccatgagat ttaccctctc tcgagatctg
     2821 cgatctgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2881 cctaactccg cccagttccg cccattctcc gccccatcgc tgactaattt tttttattta
     2941 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3001 tggaggccta ggcttttgca aaaagcttgg cattccggta ctgttggtaa agccaccatg
     3061 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
     3121 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
     3181 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
     3241 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
     3301 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
     3361 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
     3421 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
     3481 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
     3541 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     3601 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
     3661 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     3721 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     3781 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     3841 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     3901 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     3961 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     4021 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     4081 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     4141 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     4201 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     4261 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     4321 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     4381 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     4441 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     4501 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     4561 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     4621 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     4681 gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga
     4741 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4801 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4861 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     4921 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctg
     4981 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     5041 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     5101 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5161 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5221 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5281 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5341 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5401 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5461 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctc
//
