LOCUS       sequence-200668-        6204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses  E. coli RecE residues 564-868.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  AUTHORS   Zhang J, Xing X, Herr AB, Bell CE
  TITLE     Crystal structure of E. coli RecE protein reveals a toroidal 
            tetramer for processing double-stranded DNA breaks.
  JOURNAL   Structure. 2009 May 13;17(5):690-702. doi: 
            10.1016/j.str.2009.03.008.
  PUBMED    19446525
REFERENCE   2  (bases 1 to 6204)
  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"; doi: 
            "10.1016/j.str.2009.03"; journalName: "Structure"; date: 
            "2009-05-13- 13"; volume: "17"; issue: "5"; pages: "690-702"
COMMENT     Sequence Label: pET28b – E. coli RecE residues 564-868
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        64..82
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    83..107
                     /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)."
     RBS             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             163..180
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             190..207
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1132..1149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1202..1220)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1216..1263
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1300..1755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1387..1406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1597..1618
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1847..2662)
                     /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(1847..2662)
                     /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     2570..2589
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2654..2673
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2784..3372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3273..3292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3526..3543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3558..3697
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3623..3642)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3783..3805
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3799..3990)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4799..5881)
                     /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     5843..5862
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5882..5959)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6165..6184
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
       61 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
      121 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
      181 agcagcggcc tggtgccgcg cggcagccat atggaacatc cgcacaatga gaatgctggc
      241 agcgatccgc atcgcgattg cagtgatgaa actggcgaag tcgcagatcc cgtaatcgta
      301 gaagacatag agccaggtat ttattacgga atttcgaatg agaattacca cgcgggtccc
      361 ggtatcagta agtctcagct cgatgacatt gctgatactc cggcactata tttgtggcgt
      421 aaaaatgccc ccgtggacac cacaaagaca aaaacgctcg atttaggaac tgctttccac
      481 tgccgggtac ttgaaccgga agaattcagt aaccgcttta tcgtagcacc tgaatttaac
      541 cgccgtacaa acgccggaaa agaagaagag aaagcgtttc tgatggaatg cgcaagcaca
      601 ggaaaaacgg ttatcactgc ggaagaaggc cggaaaattg aactcatgta tcaaagcgtt
      661 atggctttgc cgctggggca atggcttgtt gaaagcgccg gacacgctga atcatcaatt
      721 tactgggaag atcctgaaac aggaattttg tgtcggtgcc gtccggacaa aattatccct
      781 gaatttcact ggatcatgga cgtgaaaact acggcggata ttcaacgatt caaaaccgct
      841 tattacgact accgctatca cgttcaggat gcattctaca gtgacggtta tgaagcacag
      901 tttggagtgc agccaacttt cgtttttctg gttgccagca caactattga atgcggacgt
      961 tatccggttg aaattttcat gatgggcgaa gaagcaaaac tggcaggtca acaggaatat
     1021 caccgcaatc tgcgaaccct gtctgactgc ctgaataccg atgaatggcc agctattaag
     1081 acattatcac tgccccgctg ggctaaggaa tatgcaaatg actaactcga gcaccaccac
     1141 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1201 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1261 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     1321 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1381 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1441 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1501 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1561 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1621 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1681 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1741 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1801 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     1861 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     1921 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     1981 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     2041 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     2101 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     2161 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     2221 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     2281 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     2341 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     2401 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     2461 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     2521 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     2581 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     2641 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     2701 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2761 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2821 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2881 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     2941 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3001 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3061 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3121 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3181 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3241 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3301 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3361 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3421 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3481 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3541 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3601 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3661 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3721 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3781 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     3841 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     3901 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     3961 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4021 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4081 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4141 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     4201 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4261 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     4321 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     4381 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4441 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4501 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4561 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4621 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4681 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4741 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4861 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4921 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4981 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     5041 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     5101 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5161 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     5221 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     5281 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5341 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5401 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5461 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5521 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5581 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5641 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5701 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5761 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5821 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5881 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5941 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     6001 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     6061 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     6121 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     6181 taggcgccag caaccgcacc tgtg
//
