LOCUS       sequence-244247-        4422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the hRad51(K133R) protein in mammalian cells under CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4422)
  AUTHORS   Rees HA, Yeh WH, Liu DR
  TITLE     Development of hRad51-Cas9 nickase fusions that mediate HDR without 
            double-stranded breaks.
  JOURNAL   Nat Commun. 2019 May 17;10(1):2212. doi: 10.1038/s41467-019-09983-4.
  PUBMED    31101808
REFERENCE   2  (bases 1 to 4422)
  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"; journalName: "Nat 
            Commun."; date: "2019-05-17"; volume: "10(1):2212. doi"; pages: " 
            10.1038/s41467-019-09983-4"
COMMENT     Sequence Label: hRad51(K133R)
FEATURES             Location/Qualifiers
     source          1..4422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1477..1494)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1483..1707
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1778..1794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1778..1794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1791..1813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1802..1818
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1826..1832,1833..1850,1851..1856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1871..1892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2009..2026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2180..2768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2260..2279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2939..3730,3731..3799))
                     /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     3562..3581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3800..3904)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3983..4002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(4174..4422,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggcgatgcag
      421 atgcagttgg aagcgaatgc agatactagt gtcgaggaag agtcatttgg cccgcaaccc
      481 atctcgcgtt tagagcaatg tggcatcaat gcaaacgatg tgaaaaaatt agaggaagct
      541 ggattccaca cggtcgaagc ggtcgcatac gcaccgaaaa aagagctgat caacatcaaa
      601 ggcatcagcg aggcgaaagc cgataagatt cttgcagagg cggcgaaatt agttcccatg
      661 ggatttacga cggcgactga gttccatcaa cgtcgttccg agatcattca aatcacgacc
      721 ggaagcaagg agttggataa actgcttcaa ggaggcatcg aaactggctc gatcactgaa
      781 atgtttggag agtttcgtac tggccgaacc cagatctgcc acacattggc cgtcacttgc
      841 cagcttccga tcgatcgtgg tgggggcgag gggaaggcca tgtacattga tactgagggg
      901 acgtttcgtc ccgaacgcct tcttgcggtg gcagagcgtt acggcttgtc agggtctgat
      961 gttcttgaca atgttgccta tgctcgcgca tttaataccg accaccagac gcagcttctg
     1021 taccaagcaa gcgcgatgat ggtagagtct cgttacgctt tattgattgt tgactcagca
     1081 acagcgttgt accgcacaga ttattctggg cgcggcgaat tgtcggcacg ccaaatgcat
     1141 ttagcccgct ttttgcgtat gttgctgcgc cttgcagacg aatttggggt ggctgtggtg
     1201 atcaccaacc aagttgtagc acaggtggac ggtgccgcta tgtttgccgc tgatccgaaa
     1261 aaacccatcg gagggaatat tatcgcacat gcatcgacaa ctcgcttata tctgcgtaag
     1321 ggccgtggag aaacacgcat ctgcaaaatc tatgactccc catgcctgcc ggaggcggaa
     1381 gccatgttcg caattaacgc agatggcgta ggggacgcca aagattgatg actggtacac
     1441 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     1501 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1561 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1621 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1681 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     1741 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1801 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1861 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1921 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1981 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3901 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
     3961 atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     4021 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     4081 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     4141 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     4201 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4261 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4321 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4381 atttacggta aactgcccac ttggcagtac atcaagtgta tc
//
