LOCUS       sequence-461194-        5561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near VPS13B, chr8:99439983-99440484 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5561)
  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_VPS13B
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(571..1362,1363..1431))
                     /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     1194..1213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1432..1536)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1563..2018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1650..1669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1860..1881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2149..2197
                     /note="synthetic polyadenylation signal"
     misc_feature    2211..2302
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2251..2270
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2902..3098
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2949..3084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3128..3137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3134..4786
                     /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(3200..3218)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4827..4948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4838..4857)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4892..4911
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5202..5219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5373..5561,1..400))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5453..5472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
       61 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      241 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      361 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      421 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      481 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      541 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
      601 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
      661 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
      721 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
      781 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
      841 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      901 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      961 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1021 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1081 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1141 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1201 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1261 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1321 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1381 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1441 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1501 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1561 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1621 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1681 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1741 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1801 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1861 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1921 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1981 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
     2041 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
     2101 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
     2161 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     2221 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     2281 gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtg ctagcaaacc
     2341 tcacatatag ggccaaggac ttacagaggc atatatcttt caaatagcac ataaactaga
     2401 aacttcaggg aaaaatgcat tgcagtcatt tcatttatta aaataagaat aacaaataat
     2461 ttttacttta ttggtttatc ttcagtatat gatacttaga tatgtagccc ttgacctagt
     2521 aggaatcaaa atagaggtgg ttccacttca agaatttgta ataatccctt cactttaaca
     2581 catgaatgtg ttcagtattg gcaaaagaac acgttgctac caaaactgaa acaagatcct
     2641 tgaaacagaa aggatgttgc catggaaaaa tgaagtaaca ctcagtgcag ctgtgatgca
     2701 ttcatgaata ggatattttt ttaaaagatc atttgtgttt ggtgtggctc agagggtccc
     2761 aaccagatgg tttccatgaa catgatctga cccccagctt gaggacttct tactggcaga
     2821 ttacttcatt ttctttccct tggattgtca gatccttaca ttaattcaga atttactgct
     2881 gcagactcga gatctgcgat ctgcatctca attagtcagc aaccatagtc ccgcccctaa
     2941 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catcgctgac
     3001 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3061 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttggcatt ccggtactgt
     3121 tggtaaagcc accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc
     3181 gctggaagat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt
     3241 tcctggaaca attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta
     3301 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca
     3361 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt
     3421 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa
     3481 cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat
     3541 tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac
     3601 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt
     3661 taatgaatac gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat
     3721 gaactcctct ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg
     3781 cgtgagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc
     3841 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt
     3901 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag
     3961 ccttcaggat tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc
     4021 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg
     4081 cgctcccctc tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat
     4141 caggcaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     4201 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     4261 ggataccggg aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat
     4321 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     4381 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga
     4441 ccgcctgaag tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc
     4501 catcttgctc caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga
     4561 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     4621 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     4681 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     4741 agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agagtcgggg
     4801 cggccggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     4861 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4921 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4981 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     5041 aatcgataag gatctggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     5101 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     5161 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     5221 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc t
//
