LOCUS       sequence-203161-        6640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Human RAD51 protein codon optimzed for E.coli and the E.coli
            GorE Operon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  AUTHORS   Subramanyam S, Ismail M, Bhattacharya I, Spies M
  TITLE     Tyrosine phosphorylation stimulates activity of human RAD51 
            recombinase through altered nucleoprotein filament dynamics.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Oct 11;113(41):E6045-E6054. Epub 2016
            Sep 26.
  PUBMED    27671650
REFERENCE   2  (bases 1 to 6640)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2016-10-11"; volume: "113(41)"; pages: 
            "E6045-E6054. Epub 2016 Sep 26"
COMMENT     Sequence Label: pCH1/RAD51o
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1010..1087)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1133..1151
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1152..1176
                     /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        2305..2323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2324..2348
                     /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)."
     CDS             2436..2489
                     /codon_start=1
                     /product="GroES chaperone mobile loop that interacts with 
                     GroEL"
                     /label=S loop
                     /translation="EVETKSAGGIVLTGSAAA"
     CDS             4436..4480
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4518..4536)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4532..4579
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4809..5624)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4809..5624)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5532..5551
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5625..5716)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5734..6369)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(5879..5898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6567..6640,1..1009))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
       61 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
      121 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
      181 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
      241 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
      301 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
      361 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
      421 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
      481 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
      541 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
      601 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
      661 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
      721 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
      781 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
      841 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
      901 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
      961 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     1021 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     1081 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga
     1141 ctcactatag gggaattgtg agcggataac aattcccctg tagaaataat tttgtttaac
     1201 tttaataagg agatatacca tggcaatgca gatgcagctg gaagcgaacg cagatacctc
     1261 agtggaagaa gaaagttttg gtccgcaacc gattagccgc ctggaacagt gcggtatcaa
     1321 cgcaaatgat gtgaaaaagc tggaagaagc tggctttcat accgttgaag cagtcgctta
     1381 tgcgccgaaa aaggaactga tcaacatcaa gggtatcagc gaagccaaag cagacaagat
     1441 cctggccgaa gcggccaaac tggttccgat gggctttacc acggcaaccg aatttcatca
     1501 gcgtcgcagc gaaattatcc aaattaccac gggttctaaa gaactggata agctgctgca
     1561 gggcggtatt gaaaccggct ctatcacgga aatgtttggt gaatttcgca ccggcaaaac
     1621 gcagatttgc cacaccctgg cggtgacgtg tcaactgccg atcgatcgtg gcggtggcga
     1681 gggtaaagcc atgtacattg acaccgaagg cacgtttcgt ccggaacgcc tgctggctgt
     1741 tgcggaacgc tatggtctga gcggctctga tgtgctggac aacgttgcct acgcacgtgc
     1801 tttcaatacc gatcatcaga cgcaactgct gtatcaggcg agtgccatga tggtcgaatc
     1861 ccgctacgcg ctgctgattg tggatagcgc aaccgctctg tatcgtacgg actactcagg
     1921 tcgcggcgaa ctgtcggcac gtcaaatgca cctggctcgt tttctgcgca tgctgctgcg
     1981 tctggcggac gaatttggtg ttgccgtggt tatcaccaac caggtcgtgg cgcaagtcga
     2041 tggcgcagct atgttcgcgg ccgacccgaa aaagccgatt ggtggcaata ttatcgcgca
     2101 cgccagtacc acgcgcctgt atctgcgtaa aggtcgcggc gaaacccgta tttgcaaaat
     2161 ctatgattcc ccgtgtctgc cggaagccga agccatgttt gccatcaatg ccgacggtgt
     2221 tggtgacgct aaggattaag cggccgcata atgcttaagt cgaacagaaa gtaatcgtat
     2281 tgtacacggc cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata
     2341 acaattcccc atcttagtat attagttaag tataagaagg agatatacat atgaatattc
     2401 gtccattgca tgatcgcgtg atcgtcaagc gtaaagaagt tgaaactaaa tctgctggcg
     2461 gcatcgttct gaccggctct gcagcggcta aatccacccg cggcgaagtg ctggctgtcg
     2521 gcaatggccg tatccttgaa aatggcgaag tgaagccgct ggatgtgaaa gttggcgaca
     2581 tcgttatttt caacgatggc tacggtgtga aatctgagaa gatcgacaat gaagaagtgt
     2641 tgatcatgtc cgaaagcgac attctggcaa ttgttgaagc gtaatccgcg cacgacactg
     2701 aacatacgaa tttaaggaat aaagataatg gcagctaaag acgtaaaatt cggtaacgac
     2761 gctcgtgtga aaatgctgcg cggcgtaaac gtactggcag atgcagtgaa agttaccctc
     2821 ggtccaaaag gccgtaacgt agttctggat aaatctttcg gtgcaccgac catcaccaaa
     2881 gatggtgttt ccgttgctcg tgaaatcgaa ctggaagaca agttcgaaaa tatgggtgcg
     2941 cagatggtga aagaagttgc ctctaaagca aacgacgctg caggcgacgg taccaccact
     3001 gcaaccgtac tggctcaggc tatcatcact gaaggtctga aagctgttgc tgcgggcatg
     3061 aacccgatgg acctgaaacg tggtatcgac aaagcggtta ccgttgcagt tgaagaactg
     3121 aaagcgctgt ccgtaccatg ctctgactct aaagcgattg ctcaggttgg taccatctcc
     3181 gctaactccg acgaaaccgt aggtaaactg atcgctgaag cgatggacaa agtcggtaaa
     3241 gaaggcgtta tcaccgttga agacggtacc ggtctgcagg acgaactgga cgtggttgaa
     3301 ggtatgcagt tcgaccgtgg ctacctgtct ccttacttca tcaacaagcc ggaaactggc
     3361 gcagtagaac tggaaagccc gttcatcctg ctggctgaca agaaaatctc caacatccgc
     3421 gaaatgctgc cggttctgga agctgttgcc aaagcaggca aaccgctgct gatcatcgct
     3481 gaagatgtag aaggcgaagc gctggcaact ctggttgtta acaccatgcg tggcatcgtg
     3541 aaagtcgctg cggttaaagc accgggcttc ggcgatcgtc gtaaagctat gctgcaggat
     3601 atcgcaaccc tgactggcgg taccgtgatc tctgaagaga tcggtatgga gctggaaaaa
     3661 gcaaccctgg aagacctggg tcaggctaaa cgtgttgtga tcaacaaaga caccaccact
     3721 atcatcgatg gcgtgggtga agaagctgca atccagggcc gtgttgctca gatccgtcag
     3781 cagattgaag aagcaacttc tgactacgac cgtgaaaaac tgcaggaacg cgtagcgaaa
     3841 ctggcaggcg gcgttgcagt tatcaaagtg ggtgctgcta ccgaagttga aatgaaagag
     3901 aaaaaagcac gcgttgaaga tgccctgcac gcgacccgtg ctgcggtaga agaaggcgtg
     3961 gttgctggtg gtggtgttgc gctgatccgc gtagcgtcta aactggctga cctgcgtggt
     4021 cagaacgaag accagaacgt gggtatcaaa gttgcactgc gtgcaatgga agctccgctg
     4081 cgtcagatcg tattgaactg cggcgaagaa ccgtctgttg ttgctaacac cgttaaaggc
     4141 ggcgacggca actacggtta caacgcagca accgaagaat acggcaacat gatcgacatg
     4201 ggtatcctgg atccaaccaa agtaactcgt tctgctctgc agtacgcagc ttctgtggct
     4261 ggcctgatga tcaccaccga atgcatggtt accgacctgc cgaaaaacga tgcagctgac
     4321 ttaggcgctg ctggcggtat gggcggcatg ggtggcatgg gcggcatgat gtaaagatct
     4381 caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt ctggtaaaga
     4441 aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt
     4501 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     4561 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct
     4621 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     4681 ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt cggggaaatg
     4741 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4801 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     4861 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     4921 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     4981 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5041 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     5101 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5161 cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca
     5221 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     5281 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     5341 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     5401 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     5461 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     5521 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     5581 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa
     5641 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5701 tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg ccaggaggat
     5761 acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct ccgcccccct
     5821 gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     5881 agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc cgtcctgcgg
     5941 cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt gtagacagtt
     6001 cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg cgccttatcc
     6061 ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact ggcagcagcc
     6121 attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt ggcctaacag
     6181 aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag ttaccaggtt
     6241 aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg gttttttcgt
     6301 ttacagagca ggagattacg acgatcgtaa aaggatctca agaagatcct ttacggattc
     6361 ccgacaccat cactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     6421 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
     6481 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     6541 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6601 gccagctgca ttaatgaatc ggccaacgcg cggggagagg
//
