LOCUS       sequence-466712-        5612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near RAD51B, chr14:68316358-68316859 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5612)
  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_RAD51B
FEATURES             Location/Qualifiers
     source          1..5612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(592..1383,1384..1452))
                     /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     1215..1234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1453..1557)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1584..2039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1671..1690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1881..1902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2170..2218
                     /note="synthetic polyadenylation signal"
     misc_feature    2232..2323
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2272..2291
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2974..3170
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3021..3156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3200..3209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3206..4858
                     /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(3272..3290)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4899..5020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4910..4929)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4964..4983
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5274..5291)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5445..5612,1..421))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5525..5544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
       61 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      121 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      181 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      241 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
      301 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
      361 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
      421 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      481 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      541 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      601 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
      661 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
      721 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
      781 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      841 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      901 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      961 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1021 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1081 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1141 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1201 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1261 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1321 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1381 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1441 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1501 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1561 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1621 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1681 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1741 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1801 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1861 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     1921 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     1981 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg
     2041 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2101 attacgccag cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg
     2161 agcggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     2221 tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa
     2281 taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tctatcgata ggtaccgagc
     2341 tcttacgcgt gctagcaagt tctgcatttc acctacccaa gggagtctga gagtgtgtaa
     2401 tttgggccag ggcacaacag cccttggcca ggttgaagtg gacaaacaaa tggaattggt
     2461 tccagtttac caacctggca cgaattatgt gctggtcaag gaagtgtttt tcaaatcatg
     2521 gggagccact agtgaaaatc tactggtgct ttgatggtgg cctttggaga tacttttata
     2581 atttgaaacc attcccatgg caggcccagg gagaggtctt actttagcca acatgcaaca
     2641 ttaggtagaa gaaatggggc ctttctgact ggtttgacct gtgggtgggc cctcttccac
     2701 atgtctctaa ttaggatgca agaaagagta gatattctgt ggcgactctc ctttccttct
     2761 cggctgtgag acagaacaac agcgcctttg tgatgacctt tacagggctg tttacatttg
     2821 gtgacctgcc tgaacactcc cgccagagtg gacgcacctc ttggctaatg aggaacagcc
     2881 atgctgtctt ttactatggt agaccttgga ggttaaaatg aaaaataaga ggatcctctg
     2941 ttgatgacta gtcgaagctc gagatctgcg atctgcatct caattagtca gcaaccatag
     3001 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3061 cccatcgctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3121 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttggca
     3181 ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc
     3241 gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag
     3301 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac
     3361 ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
     3421 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
     3481 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
     3541 tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg
     3601 gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat
     3661 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
     3721 acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat
     3781 tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca
     3841 tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat
     3901 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
     3961 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     4021 gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt
     4081 ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     4141 tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca
     4201 tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac
     4261 acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     4321 ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt
     4381 gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     4441 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     4501 cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc
     4561 tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct
     4621 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     4681 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     4741 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     4801 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt
     4861 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     4921 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4981 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     5041 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     5101 caaatgtggt aaaatcgata aggatctgga tccgtcgacc gatgcccttg agagccttca
     5161 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     5221 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
     5281 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5341 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5401 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5461 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5521 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5581 ctgccgctta ccggatacct gtccgccttt ct
//
