LOCUS       sequence-466686-        5914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  chr8:16099746-16100677 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5914)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5914)
  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_MSR1
FEATURES             Location/Qualifiers
     source          1..5914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        920..1116
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      967..1102
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1146..1155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1152..2804
                     /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(1218..1236)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2845..2966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2856..2875)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2910..2929
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3220..3237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3391..3979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3471..3490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4150..4941,4942..5010))
                     /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     4773..4792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5011..5115)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5142..5597
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5229..5248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5439..5460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5728..5776
                     /note="synthetic polyadenylation signal"
     misc_feature    5790..5881
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5830..5849
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gccacatcca gagtcatggg ccattgttct gggggagagt cctaccaaac taaaactaag
       61 atacctcctt tattccttcc agctgacagg acatgagccg tatggtagct ctcttccaag
      121 accccgcagc ctggggtaat aaattctgcc aagctctccc tctttctctc tctgctattc
      181 cctgaagtct gacaaccctg cgggtcagcc ccctagggcc atccagcctc cgtcgcccga
      241 tgccaatttt accttgtgac tctcacaaca gggggaaaat ttagcattcc ttggagaccg
      301 aatgggatgc cgtgagctta agcccttcca agagcttagc aaccagtctg tccttattcc
      361 gtcccaataa cacagccccc gatggaacaa ttacaaaagt atttacaacg ccttacctct
      421 tactgttctt atagatgtac ttattcctgt aggtagactc taaaaagttt gaagaggaaa
      481 aattatgaaa atcaagaggg agaagttgtt ggaaacagcg agttcctctt ctgaggttac
      541 acttgtcctc tgttttcaaa gcctaacttc cttgcctcct tgcccctagt taggataaac
      601 aaccttccag ccatttccaa tctgtaaccc acatccatta acccacatct gttcccaatc
      661 tgtaacccac atccattccc aatctgtaac aactcacatc tgttcctgat ttggtaccct
      721 tagttccgaa gttgctctgt agcccccacc cctgctccat ttgaagtagt caatgaggat
      781 cagcttagat tgtgtcgtcg gaccccaacc tatggagact ggacacagta gcagggactg
      841 actatgttag agacacaaat ccttcccttc tttgttctgt gcactctcgc cattgctccc
      901 tccctcgaga tctgcgatct gcatctcaat tagtcagcaa ccatagtccc gcccctaact
      961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tcgctgacta
     1021 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     1081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttggcattcc ggtactgttg
     1141 gtaaagccac catggaagac gccaaaaaca taaagaaagg cccggcgcca ttctatccgc
     1201 tggaagatgg aaccgctgga gagcaactgc ataaggctat gaagagatac gccctggttc
     1261 ctggaacaat tgcttttaca gatgcacata tcgaggtgga catcacttac gctgagtact
     1321 tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat acaaatcaca
     1381 gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg ggcgcgttat
     1441 ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca
     1501 gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa aaaggggttg caaaaaattt
     1561 tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat tatcatggat tctaaaacgg
     1621 attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct cccggtttta
     1681 atgaatacga ttttgtgcca gagtccttcg atagggacaa gacaattgca ctgatcatga
     1741 actcctctgg atctactggt ctgcctaaag gtgtcgctct gcctcataga actgcctgcg
     1801 tgagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg gatactgcga
     1861 ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc ggatatttga
     1921 tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt ctgaggagcc
     1981 ttcaggatta caagattcaa agtgcgctgc tggtgccaac cctattctcc ttcttcgcca
     2041 aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct tctggtggcg
     2101 ctcccctctc taaggaagtc ggggaagcgg ttgccaagag gttccatctg ccaggtatca
     2161 ggcaaggata tgggctcact gagactacat cagctattct gattacaccc gagggggatg
     2221 ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt gtggatctgg
     2281 ataccgggaa aacgctgggc gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga
     2341 ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac aaggatggat
     2401 ggctacattc tggagacata gcttactggg acgaagacga acacttcttc atcgttgacc
     2461 gcctgaagtc tctgattaag tacaaaggct atcaggtggc tcccgctgaa ttggaatcca
     2521 tcttgctcca acaccccaac atcttcgacg caggtgtcgc aggtcttccc gacgatgacg
     2581 ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg acggaaaaag
     2641 agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg
     2701 tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga aaaatcagag
     2761 agatcctcat aaaggccaag aagggcggaa agatcgccgt gtaattctag agtcggggcg
     2821 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     2881 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2941 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3001 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3061 tcgataagga tctggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     3121 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     3181 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     3241 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3301 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3361 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3421 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3481 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3541 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3601 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3661 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3721 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3781 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3841 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3901 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3961 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4021 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4081 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4141 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4201 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4261 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4321 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4381 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4441 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4501 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4561 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4621 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4681 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4741 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4801 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4861 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4921 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4981 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5041 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5101 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     5161 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5221 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5281 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5341 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5401 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5461 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5521 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5581 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5641 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     5701 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     5761 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     5821 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     5881 tatcgatagg taccgagctc ttacgcgtgc tagc
//
