LOCUS       sequence-455992-        8557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in BL21(DE3) of T4 SbcC-like subunit of palindrome 
            specific endonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8557)
  AUTHORS   Herdendorf TJ, Albrecht DW, Benkovic SJ, Nelson SW
  TITLE     Biochemical characterization of bacteriophage T4 Mre11-Rad50 
            complex.
  JOURNAL   J Biol Chem. 2011 Jan 28;286(4):2382-92. doi: 
            10.1074/jbc.M110.178871. Epub 2010 Nov 15.
  PUBMED    21081488
REFERENCE   2  (bases 1 to 8557)
  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: 
            "10.1074/jbc.M110.178871"; journalName: "J Biol Chem"; date: 
            "2011-01-28- 28"; volume: "286"; issue: "4"; pages: "2382-92"
COMMENT     Sequence Label: pTYB1-T4gp46
FEATURES             Location/Qualifiers
     source          1..8557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(286..799)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(481..502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     693..712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      910..1498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1399..1418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1652..1669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1691..1710)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1851..1873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1867..2058)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2615..3697)
                     /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     3659..3678
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3698..3775)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      3928..4014
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4111..4197
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4332..4351
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4332..4350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4351..4375
                     /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             4390..4412
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6100..6918
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae VMA1"
                     /product="5' region of the intein from the yeast Vma1 
                     subunit of the vacuolar ATPase (Chong et al., 1998)"
                     /label=Sce VMA intein 5' region
                     /translation="CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRE
                     TMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEV
                     ITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARD
                     LSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSD
                     RATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG"
     CDS             7374..7385
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7495..7650
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(7715..7733)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7729..7776
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        7836..7940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7941..8009,8010..8557,1..244)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8159..8178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
       61 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
      121 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      181 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
      241 gtaactgtca gaccaagttt actcatatat actttagatt gatttacccc ggttgataat
      301 cagaaaagcc ccaaaaacag gaagattgta taagcaaata tttaaattgt aaacgttaat
      361 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      421 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagggtt gagtgttgtt
      481 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
      541 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
      601 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
      661 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
      721 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
      781 gcgccgctac agggcgcgta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      841 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      901 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      961 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1021 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1081 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1141 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1201 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1261 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1321 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1381 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1441 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1501 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1561 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1621 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagctatggt
     1681 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     1741 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     1801 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     1861 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     1921 atcagcgtgg tcgtgcagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     1981 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     2041 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     2101 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     2161 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     2221 aatcactcag ggtcaatgcc agccgaacgc cagcaagacg tagcccagcg cgtcggccgc
     2281 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     2341 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     2401 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     2461 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     2521 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     2581 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2641 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2701 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     2761 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     2821 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     2881 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     2941 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3001 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3061 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     3121 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     3181 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     3241 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     3301 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     3361 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3421 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3481 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3541 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3601 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     3661 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     3721 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     3781 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     3841 ccgttgagca ccgccgccgc aaggaatggt gcatgccggc atgccgccct ttcgtcttca
     3901 agaattaatt cccaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3961 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     4021 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     4081 taaactgcca ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac
     4141 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4201 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     4261 ccataaactg ccaggaattg gggatcggaa ttaattcccg gtttaaaccg gggatctcga
     4321 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     4381 agaaataatt ttgtttaact ttaagaagga gatatacata tgaagaattt taaacttaat
     4441 agagtaaaat ataaaaatat aatgtcagtt ggtcaaaatg ggattgacat tcaattagat
     4501 aaggttcaaa aaactcttat tactggacga aatggcggtg gtaagtctac tatgctagaa
     4561 gccatcacat ttgggctttt tggtaagcca tttcgcgatg taaagaaagg tcaattaata
     4621 aacagcacaa ataagaaaga acttttagtt gaactgtgga tggaatatga tgagaaaaag
     4681 tactatatca aaagaggaca aaaaccaaac gttttcgaaa tcaccgttaa cggtacacgt
     4741 cttaatgaat ctgccagcag taaagatttc caagcagaat ttgaacagct tatcggaatg
     4801 tcatacgcca gtttcaagca gattgttgtc cttggtacag cagggtatat ccctttcatg
     4861 ggtttgtcga cccctgcgcg aagaaagctt gtggaagacc tgcttgaggt aggaacatta
     4921 gctgaaatgg ataagcttaa taaagcacta atacgtgaat taaattcgca gaaccaagtg
     4981 cttgatgtta aaaaagatag tattatccaa caaattaaaa tatataatga taatgttgaa
     5041 cgccagaaaa aattaacggg tgacaacctt actcgtctac aaaatatgta tgatgatttg
     5101 gcaaaagaag ctagaacgct aaaatcggaa atagaagaag ctaacgaaag attagttaat
     5161 attgttttgg acgaagaccc gactgatgca tttaataaaa tcggtcaaga agcattttta
     5221 attaaatcaa aaattgactc gtataataaa gtcattaata tgtatcacga aggtggatta
     5281 tgtccaacct gtttgtcaca attgagttcc ggtgataaag ttgtttctaa aattaaagat
     5341 aaagtttctg aatgcacaca ttcgtttgaa cagctttcaa cacatcgtga taatttaaaa
     5401 gttcttgttg atgaataccg agataatatt aaaacccagc agtcgttggc aaatgatatt
     5461 cgcaataaaa agcaatctct gatcgcagca gtagataaag ctaaaaaagt taaagcggct
     5521 atagaaaaag catcttctga gtttattgac catgctgatg aaatagcact gcttcaagaa
     5581 gaacttgata aaattgttaa gacaaaaact aatttagtaa tggaaaaata ccaccgagga
     5641 attttgactg atatgctcaa agattctggt attaaaggtg ctattattaa aaagtacatt
     5701 ccattattta ataagcagat taaccattat cttaaaataa tggaagcgga ttatgtgttt
     5761 acattagatg aagaatttaa tgagacaatt aaatcccgtg gtcgtgaaga ttttagttat
     5821 gcttcattta gtgaaggtga aaaggcacga atcgatattg ctcttttatt tacttggcgt
     5881 gatattgctt ctatcgtatc tggtgttagt attagtacat taattcttga tgaagtgttt
     5941 gatgggtcat ttgatgccga aggtattaaa ggtgtagcta atattataaa ttcaatgaaa
     6001 aacactaacg tttttataat ttcgcataaa gaccatgacc cgcaagaata tggtcagcat
     6061 cttcaaatga agaaagttgg tcgatttact gtaatggcgt gctttgccaa gggtaccaat
     6121 gttttaatgg cggatgggtc tattgaatgt attgaaaaca ttgaggttgg taataaggtc
     6181 atgggtaaag atggcagacc tcgtgaggta attaaattgc ccagaggaag agaaactatg
     6241 tacagcgtcg tgcagaaaag tcagcacaga gcccacaaaa gtgactcaag tcgtgaagtg
     6301 ccagaattac tcaagtttac gtgtaatgcg acccatgagt tggttgttag aacacctcgt
     6361 agtgtccgcc gtttgtctcg taccattaag ggtgtcgaat attttgaagt tattactttt
     6421 gagatgggcc aaaagaaagc ccccgacggt agaattgttg agcttgtcaa ggaagtttca
     6481 aagagctacc caatatctga ggggcctgag agagccaacg aattagtaga atcctataga
     6541 aaggcttcaa ataaagctta ttttgagtgg actattgagg ccagagatct ttctctgttg
     6601 ggttcccatg ttcgtaaagc tacctaccag acttacgctc caattcttta tgagaatgac
     6661 cactttttcg actacatgca aaaaagtaag tttcatctca ccattgaagg tccaaaagta
     6721 cttgcttatt tacttggttt atggattggt gatggattgt ctgacagggc aactttttcg
     6781 gttgattcca gagatacttc tttgatggaa cgtgttactg aatatgctga aaagttgaat
     6841 ttgtgcgccg agtataagga cagaaaagaa ccacaagttg ccaaaactgt taatttgtac
     6901 tctaaagttg tcagaggtaa tggtattcgc aataatctta atactgagaa tccattatgg
     6961 gacgctattg ttggcttagg attcttgaag gacggtgtca aaaatattcc ttctttcttg
     7021 tctacggaca atatcggtac tcgtgaaaca tttcttgctg gtctaattga ttctgatggc
     7081 tatgttactg atgagcatgg tattaaagca acaataaaga caattcatac ttctgtcaga
     7141 gatggtttgg tttcccttgc tcgttcttta ggcttagtag tctcggttaa cgcagaacct
     7201 gctaaggttg acatgaatgt caccaaacat aaaattagtt atgctattta tatgtctggt
     7261 ggagatgttt tgcttaacgt tctttcgaag tgtgccggct ctaaaaaatt caggcctgct
     7321 cccgccgctg cttttgcacg tgagtgccgc ggattttatt tcgagttaca agaattgaag
     7381 gaagacgatt attatgggat tactttatct gatgattctg atcatcagtt tttgcttgga
     7441 tcccaggttg tcgtccatgc atgcggtggc ctgaccggtc tgaactcagg cctcacgaca
     7501 aatcctggtg tatccgcttg gcaggtcaac acagcttata ctgcgggaca attggtcaca
     7561 tataacggca agacgtataa atgtttgcag ccccacacct ccttggcagg atgggaacca
     7621 tccaacgttc ctgccttgtg gcagcttcaa tgactgcagg aaggggatcc ggctgctaac
     7681 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     7741 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     7801 taactacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7861 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     7921 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     7981 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     8041 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     8101 tccttgagag ttttcgcccc gaagaacgtt tcccaatgat gagcactttt aaagttctgc
     8161 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     8221 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     8281 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     8341 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     8401 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     8461 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     8521 gcgaactact tactctagct tcccggcaac aattaat
//
