LOCUS       sequence-463060-        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near DDX25, chr11:125935511-125936012 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5571)
  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_DDX25
FEATURES             Location/Qualifiers
     source          1..5571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    587..678
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(619..638)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    692..740
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(871..1326)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1008..1029)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1220..1239
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1353..1457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1458..1526,1527..2318)
                     /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(1676..1695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2489..3077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2978..2997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3231..3248
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3502..3623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3539..3558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3593..3612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3664..5316)
                     /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     5232..5250
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      5313..5322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5352..5548)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5366..5501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttaagatct cccagagtcg tccaactgca gagcactctc caggctggtc caggcctagg
       61 gctccgggca ggggctggtg ttgaactaag cttaccggct ttgccagccc taggtggcaa
      121 agcacccttt aaacagactt ctgccctggt gttctccgtc tcgactctcc tcccccaaac
      181 gctaggcctc cacaggttct ggcacagcca gccaagtccc acagctgttt ctaatattct
      241 gcacacactt tattgaaatg agaagccccg tttatcatcc ttcaggcccc tgctcctcct
      301 gattaggtac agatgacctg acagtcagtg gggcctgagc cagcaatcaa agaggagatt
      361 gttctaagcc aggaggtgag aggggcagag gcctgggaag ccgagcaggg tccaaggaga
      421 cttgggccag ccaggaggag atgggggagg ttctcaggct gcaggaagga ggggagggca
      481 gggctgagta aattcttcaa ggacccctgg tttgctcatc ttctctggcc atatattctc
      541 tctggcctcc gaggctagca cgcgtaagag ctcggtacct atcgatagag aaatgttctg
      601 gcacctgcac ttgcactggg gacagcctat tttgctagtt tgttttgttt cgttttgttt
      661 tgatggagag cgtatgttag tactatcgat tcacacaaaa aaccaacaca cagatgtaat
      721 gaaaataaag atattttatt gcggccgctc caagtacctc ccgtacctta atattactta
      781 cttatcatgg tagcttgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
      841 aacagttgcg cagcctgaat ggcgaatggc aaattgtaag cgttaatatt ttgttaaaat
      901 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
      961 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1021 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1081 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1141 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1201 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1261 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1321 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1441 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1501 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1561 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1621 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1681 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1741 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1801 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1861 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1921 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1981 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2041 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2101 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2161 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2221 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2281 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2341 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2401 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2461 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2521 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2581 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2641 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2701 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2761 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2821 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2881 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2941 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3001 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3061 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3121 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3181 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3241 tgagcgagga agcggaagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     3301 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     3361 ttgaaggctc tcaagggcat cggtcgacgg atccagatcc ttatcgattt taccacattt
     3421 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3481 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3541 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3601 tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc ggccggccgc cccgactcta
     3661 gaattacacg gcgatctttc cgcccttctt ggcctttatg aggatctctc tgatttttct
     3721 tgcgtcgagt tttccggtaa gacctttcgg tacttcgtcc acaaacacaa ctcctccgcg
     3781 caactttttc gcggttgtta cttgactggc gacgtaatcc acgatctctt tttccgtcat
     3841 cgtctttccg tgctccaaaa caacaacggc ggcgggaagt tcaccggcgt catcgtcggg
     3901 aagacctgcg acacctgcgt cgaagatgtt ggggtgttgg agcaagatgg attccaattc
     3961 agcgggagcc acctgatagc ctttgtactt aatcagagac ttcaggcggt caacgatgaa
     4021 gaagtgttcg tcttcgtccc agtaagctat gtctccagaa tgtagccatc catccttgtc
     4081 aatcaaggcg ttggtcgctt ccggattgtt tacataaccg gacataatca taggacctct
     4141 cacacacagt tcgcctcttt gattaacgcc cagcgttttc ccggtatcca gatccacaac
     4201 cttcgcttca aaaaatggaa caactttacc gaccgcgccc ggtttatcat ccccctcggg
     4261 tgtaatcaga atagctgatg tagtctcagt gagcccatat ccttgcctga tacctggcag
     4321 atggaacctc ttggcaaccg cttccccgac ttccttagag aggggagcgc caccagaagc
     4381 aatttcgtgt aaattagata aatcgtattt gtcaatcaga gtgcttttgg cgaagaagga
     4441 gaatagggtt ggcaccagca gcgcactttg aatcttgtaa tcctgaaggc tcctcagaaa
     4501 cagctcttct tcaaatctat acattaagac gactcgaaat ccacatatca aatatccgag
     4561 tgtagtaaac attccaaaac cgtgatggaa tggaacaaca cttaaaatcg cagtatccgg
     4621 aatgatttga ttgccaaaaa taggatctct ggcatgcgag aatctcacgc aggcagttct
     4681 atgaggcaga gcgacacctt taggcagacc agtagatcca gaggagttca tgatcagtgc
     4741 aattgtcttg tccctatcga aggactctgg cacaaaatcg tattcattaa aaccgggagg
     4801 tagatgagat gtgacgaacg tgtacatcga ctgaaatccc tggtaatccg ttttagaatc
     4861 catgataata attttttgga tgattgggag ctttttttgc acgttcaaaa ttttttgcaa
     4921 cccctttttg gaaacgaaca ccacggtagg ctgcgaaatg cccatactgt tgagcaattc
     4981 acgttcatta taaatgtcgt tcgcgggcgc aactgcaact ccgataaata acgcgcccaa
     5041 caccggcata aagaattgaa gagagttttc actgcatacg acgattctgt gatttgtatt
     5101 cagcccatat cgtttcatag cttctgccaa ccgaacggac atttcgaagt actcagcgta
     5161 agtgatgtcc acctcgatat gtgcatctgt aaaagcaatt gttccaggaa ccagggcgta
     5221 tctcttcata gccttatgca gttgctctcc agcggttcca tcttccagcg gatagaatgg
     5281 cgccgggcct ttctttatgt ttttggcgtc ttccatggtg gctttaccaa cagtaccgga
     5341 atgccaagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     5401 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagcgatgg
     5461 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     5521 actatggttg ctgactaatt gagatgcaga tcgcagatct cgagcccggt g
//
