LOCUS       sequence-7377-ex        5857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5857)
  AUTHORS   Lee BI, Kim KH, Park SJ, Eom SH, Song HK, Suh SW
  TITLE     Ring-shaped architecture of RecR: implications for its role in 
            homologous recombinational DNA repair.
  JOURNAL   EMBO J. 2004 May 19. 23(10):2029-38.
  PUBMED    15116069
REFERENCE   2  (bases 1 to 5857)
  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: "EMBO J. 
            2004 May 19. 23(10):2029-38."
COMMENT     Sequence Label: pET-28b(+)-DR RecR(34-220)
FEATURES             Location/Qualifiers
     source          1..5857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(731..748)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(758..775)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             794..816
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    831..855
                     /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)."
     primer_bind     complement(855..874)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(856..874)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(958..977)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1183..1260
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1261..2343
                     /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"
     primer_bind     complement(1280..1299)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3152..3343
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3337..3359)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3445..3587
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3500..3519
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3602..3619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3773..4361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3853..3872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4472..4491)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4483..5298
                     /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             4483..5298
                     /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     complement(4556..4575)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5391..5846)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5528..5549)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5740..5759
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gaggcgggat gcgggcaccg
      181 ctgctccgtc ggcgccgtct tcgtcaccgg gacgttgcgg cggctggggc ttgctcacgg
      241 tctggcggcc tgtgagcgca cggcccagcg tcacctcgtc ggtgtattcc agcgagccgc
      301 ccaccggcac cccgtaggcg atgcggctga tggcggcgcc gagcggttcg agcagccgct
      361 gaaggtagag cgcggtggcg tcgccctcca ccgttgtgcc agtcgccaga atcacttcca
      421 tgccttgccc cacgcggggc agcagcggct tgatgtgcag cttgtcgggg ccgacgccgt
      481 tcatcgggct cagaacgccg tgcaggacgt ggtagagccc ccggtactcg ccgctgcgct
      541 ccagcgcaat cacgtcgccg ggttcttcca ccacgcagat ggtgcgctgg tcgcggctgg
      601 ggtcggcgca cacgtcgcac ttctcggcgt cggtgatgtt gaaacaaatc gggcagacgt
      661 gcaggtcgcg cttggcttcg agcagggcgg aggcgaggcg ctcgatgtcc tcacgcggct
      721 gctccatatg gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc
      781 tgcccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta
      841 tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc
      901 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
      961 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     1021 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
     1081 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     1141 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc
     1201 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     1261 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     1321 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     1381 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     1441 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     1501 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     1561 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     1621 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     1681 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     1741 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     1801 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     1861 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     1921 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     1981 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     2041 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     2101 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     2161 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     2221 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     2281 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     2341 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat
     2401 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     2461 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     2521 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     2581 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     2641 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat
     2701 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     2761 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     2821 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     2881 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     2941 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     3001 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     3061 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     3121 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     3181 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     3241 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     3301 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3361 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3421 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     3481 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc
     3541 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3781 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3841 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3901 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3961 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4021 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4081 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4141 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4201 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4261 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4321 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4381 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaaca
     4441 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     4501 gaaacgtctt gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     4561 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     4621 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     4681 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     4741 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca
     4801 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     4861 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     4921 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     4981 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     5041 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     5101 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     5161 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     5221 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     5281 ctcgatgagt ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa
     5341 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa
     5401 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5461 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5521 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5581 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5641 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5701 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5761 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5821 gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
//
