LOCUS       sequence-466690-        5708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near MECOM, chr3:169362285-169362786 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5708)
  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_MECOM
FEATURES             Location/Qualifiers
     source          1..5708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    733..824
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(765..784)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    838..886
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(1017..1472)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1154..1175)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1366..1385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1499..1603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1604..1672,1673..2464)
                     /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     complement(1822..1841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2635..3223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3124..3143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3377..3394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3648..3769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3685..3704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3739..3758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3810..5462)
                     /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     5378..5396
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      5459..5468
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5498..5694)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5512..5647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agccccaaga ataaaggggc ccttgtgtat ggtgggcttt tcctcactgt tattcttggt
       61 ctcaacttga tgaagtgtat tcgtgatata cacgaagcac tgtgactctt gtgtctctct
      121 ttgtgagact gtccatgcct ttgttcctgc tgtcagtcag ttgttctttc tcttgggcag
      181 ccacattggt aaccgcagta tctacttgca aatagatttc tgtaaaacca tattatcctt
      241 caggccttta ctcctagccc ccacccctct ttcctcaaca atctggccat tcatgtggca
      301 tagctccctt gcttgcttct ttgccatatg tctagtggtt tgtgtttcag ggaggctgtg
      361 tgatacactg gaatgaaaac tgggctgggt gccaggaacc ttgaattttt gcacaagtca
      421 gacttgggaa cttggaaaag tcatttgact ttaccctctg aagtctcact caggtctctt
      481 gtagctgtaa ctctatgagt cttcgaattc tgtctcttac acctgctctt tagttctttt
      541 catctttatg gacatgttct atattgtcat ggttttggag gttttaattt attgttgttg
      601 ttgagaaaag cccaccaaca acaataaaca aaacctccaa aataattact tttaatggca
      661 aaactgcaat tacttttaca ccaacctaat atattggggg ctagcacgcg taagagctcg
      721 gtacctatcg atagagaaat gttctggcac ctgcacttgc actggggaca gcctattttg
      781 ctagtttgtt ttgtttcgtt ttgttttgat ggagagcgta tgttagtact atcgattcac
      841 acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg ccgctccaag
      901 tacctcccgt accttaatat tacttactta tcatggtagc ttgggctggc gtaatagcga
      961 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcaaat
     1021 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1081 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1141 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1201 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1261 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1321 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1381 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     1441 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     1501 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1561 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1621 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1681 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1741 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1801 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     1861 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1921 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1981 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2041 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2101 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2161 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2221 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2281 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2341 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2401 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2461 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2521 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2581 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2761 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2881 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2941 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3001 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3061 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3241 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3301 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3361 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgct gccggcacct
     3421 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     3481 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgacggatcc
     3541 agatccttat cgattttacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3601 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct
     3781 cgaagcggcc ggccgccccg actctagaat tacacggcga tctttccgcc cttcttggcc
     3841 tttatgagga tctctctgat ttttcttgcg tcgagttttc cggtaagacc tttcggtact
     3901 tcgtccacaa acacaactcc tccgcgcaac tttttcgcgg ttgttacttg actggcgacg
     3961 taatccacga tctctttttc cgtcatcgtc tttccgtgct ccaaaacaac aacggcggcg
     4021 ggaagttcac cggcgtcatc gtcgggaaga cctgcgacac ctgcgtcgaa gatgttgggg
     4081 tgttggagca agatggattc caattcagcg ggagccacct gatagccttt gtacttaatc
     4141 agagacttca ggcggtcaac gatgaagaag tgttcgtctt cgtcccagta agctatgtct
     4201 ccagaatgta gccatccatc cttgtcaatc aaggcgttgg tcgcttccgg attgtttaca
     4261 taaccggaca taatcatagg acctctcaca cacagttcgc ctctttgatt aacgcccagc
     4321 gttttcccgg tatccagatc cacaaccttc gcttcaaaaa atggaacaac tttaccgacc
     4381 gcgcccggtt tatcatcccc ctcgggtgta atcagaatag ctgatgtagt ctcagtgagc
     4441 ccatatcctt gcctgatacc tggcagatgg aacctcttgg caaccgcttc cccgacttcc
     4501 ttagagaggg gagcgccacc agaagcaatt tcgtgtaaat tagataaatc gtatttgtca
     4561 atcagagtgc ttttggcgaa gaaggagaat agggttggca ccagcagcgc actttgaatc
     4621 ttgtaatcct gaaggctcct cagaaacagc tcttcttcaa atctatacat taagacgact
     4681 cgaaatccac atatcaaata tccgagtgta gtaaacattc caaaaccgtg atggaatgga
     4741 acaacactta aaatcgcagt atccggaatg atttgattgc caaaaatagg atctctggca
     4801 tgcgagaatc tcacgcaggc agttctatga ggcagagcga cacctttagg cagaccagta
     4861 gatccagagg agttcatgat cagtgcaatt gtcttgtccc tatcgaagga ctctggcaca
     4921 aaatcgtatt cattaaaacc gggaggtaga tgagatgtga cgaacgtgta catcgactga
     4981 aatccctggt aatccgtttt agaatccatg ataataattt tttggatgat tgggagcttt
     5041 ttttgcacgt tcaaaatttt ttgcaacccc tttttggaaa cgaacaccac ggtaggctgc
     5101 gaaatgccca tactgttgag caattcacgt tcattataaa tgtcgttcgc gggcgcaact
     5161 gcaactccga taaataacgc gcccaacacc ggcataaaga attgaagaga gttttcactg
     5221 catacgacga ttctgtgatt tgtattcagc ccatatcgtt tcatagcttc tgccaaccga
     5281 acggacattt cgaagtactc agcgtaagtg atgtccacct cgatatgtgc atctgtaaaa
     5341 gcaattgttc caggaaccag ggcgtatctc ttcatagcct tatgcagttg ctctccagcg
     5401 gttccatctt ccagcggata gaatggcgcc gggcctttct ttatgttttt ggcgtcttcc
     5461 atggtggctt taccaacagt accggaatgc caagcttttt gcaaaagcct aggcctccaa
     5521 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     5581 aaataaaaaa aattagtcag cgatggggcg gagaatgggc ggaactgggc ggagttaggg
     5641 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcagatcgc
     5701 agatctcg
//
