LOCUS       sequence-455993-        7894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in BL21(DE3) of T4 SbcD-like subunit of palindrome 
            specific endonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7894)
  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 7894)
  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-T4gp47
FEATURES             Location/Qualifiers
     source          1..7894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     402..421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(640..744)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(804..851)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     847..865
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(930..1085)
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     CDS             complement(1195..1206)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(1662..2480)
                     /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"
     RBS             3505..3527
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3542..3566
                     /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(3566..3585)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3567..3585)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(3720..3806)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3903..3989)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        4142..4219
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4220..5302
                     /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(4239..4258)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5859..6050
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6044..6066)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6207..6226
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6248..6265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6419..7007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6499..6518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      7118..7631
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7205..7224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7415..7436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(7673..7894,1..570,571..639))
                     /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"
ORIGIN
        1 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
       61 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      121 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      181 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
      241 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
      301 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      361 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
      421 cagaacttta aaagtgctca tcattgggaa acgttcttcg gggcgaaaac tctcaaggat
      481 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
      541 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
      601 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      661 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      721 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtagttat
      781 ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
      841 ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg
      901 ttagcagccg gatccccttc ctgcagtcat tgaagctgcc acaaggcagg aacgttggat
      961 ggttcccatc ctgccaagga ggtgtggggc tgcaaacatt tatacgtctt gccgttatat
     1021 gtgaccaatt gtcccgcagt ataagctgtg ttgacctgcc aagcggatac accaggattt
     1081 gtcgtgaggc ctgagttcag accggtcagg ccaccgcatg catggacgac aacctgggat
     1141 ccaagcaaaa actgatgatc agaatcatca gataaagtaa tcccataata atcgtcttcc
     1201 ttcaattctt gtaactcgaa ataaaatccg cggcactcac gtgcaaaagc agcggcggga
     1261 gcaggcctga attttttaga gccggcacac ttcgaaagaa cgttaagcaa aacatctcca
     1321 ccagacatat aaatagcata actaatttta tgtttggtga cattcatgtc aaccttagca
     1381 ggttctgcgt taaccgagac tactaagcct aaagaacgag caagggaaac caaaccatct
     1441 ctgacagaag tatgaattgt ctttattgtt gctttaatac catgctcatc agtaacatag
     1501 ccatcagaat caattagacc agcaagaaat gtttcacgag taccgatatt gtccgtagac
     1561 aagaaagaag gaatattttt gacaccgtcc ttcaagaatc ctaagccaac aatagcgtcc
     1621 cataatggat tctcagtatt aagattattg cgaataccat tacctctgac aactttagag
     1681 tacaaattaa cagttttggc aacttgtggt tcttttctgt ccttatactc ggcgcacaaa
     1741 ttcaactttt cagcatattc agtaacacgt tccatcaaag aagtatctct ggaatcaacc
     1801 gaaaaagttg ccctgtcaga caatccatca ccaatccata aaccaagtaa ataagcaagt
     1861 acttttggac cttcaatggt gagatgaaac ttactttttt gcatgtagtc gaaaaagtgg
     1921 tcattctcat aaagaattgg agcgtaagtc tggtaggtag ctttacgaac atgggaaccc
     1981 aacagagaaa gatctctggc ctcaatagtc cactcaaaat aagctttatt tgaagccttt
     2041 ctataggatt ctactaattc gttggctctc tcaggcccct cagatattgg gtagctcttt
     2101 gaaacttcct tgacaagctc aacaattcta ccgtcggggg ctttcttttg gcccatctca
     2161 aaagtaataa cttcaaaata ttcgacaccc ttaatggtac gagacaaacg gcggacacta
     2221 cgaggtgttc taacaaccaa ctcatgggtc gcattacacg taaacttgag taattctggc
     2281 acttcacgac ttgagtcact tttgtgggct ctgtgctgac ttttctgcac gacgctgtac
     2341 atagtttctc ttcctctggg caatttaatt acctcacgag gtctgccatc tttacccatg
     2401 accttattac caacctcaat gttttcaata cattcaatag acccatccgc cattaaaaca
     2461 ttggtaccct tggcaaagca cgctgttgcc tctacatata gctgatttgc atattgaata
     2521 agtgcttcac ggtcagaatc agtgatgtct ggaattgcat taatatactc ttccattaat
     2581 gtctgaagcg actgaacttc aacttcttca ctgtcatctg actcgacaga gttatcaatc
     2641 tttgacacaa ctcgtaatga atgcacaact ttttctagtt cagattcgaa cttcgtcaga
     2701 tttttgtcta cttcagttac tataacacgt actgatagat ttgtaaaatc tttatagtca
     2761 atttttcctt taaatggata atgaattcta cgatgccagg tagtattgtt tggaataaat
     2821 tccgttcgtt ctgtttctgt atcaaacatc cagaacccac gagggtcatt ctcgtcacct
     2881 gcggttagtg tccatggtgt cccaatatat ctgacgtttg cagcctcaga aatagtatgg
     2941 aagtgaccag accacacttc tttataagtc ttaaggaaat cgggttcaag accatgagat
     3001 ttcattcctt tataaaaata aaatccattc agttcccagt gaccaacaca aaaagaagca
     3061 gatgaagttt tgatatgctc aagaatttca ccagtatttt cttcgcacat ccaaggaatc
     3121 aaatcaatca aacacccgtc aaaatctact gtagtaggct tatcatacac tttaacatta
     3181 ggatatttag ccaaaagctc agtagaagca tttggatgca ttacattttt atagtggaga
     3241 tcgtgatttc ctacaatagt gtgtaatgta attccagcat catcaagcgt ttgaactatt
     3301 tcacgggcaa actccatagt tttatgtgtg atcgcttttc gcacatcaaa aatatcaccg
     3361 tattgaatcc aggtagtaat tccatttttc ttagaatatt ctatcgcttg cttaattcca
     3421 tcaatttgaa taccgcgaat ccactcatca tcagctttaa cgcctaaatg ccaatcacct
     3481 aaatttaaaa ttttcatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag
     3541 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
     3601 gatccccggt ttaaaccggg aattaattcc gatccccaat tcctggcagt ttatggcggg
     3661 cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga
     3721 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3781 ttcgactgag cctttcgttt tatttgatgc ctggggaatt aattcctggc agtttatggc
     3841 gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc
     3901 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     3961 tctttcgact gagcctttcg ttttatttga tgcctgggga attgggaatt aattcttgaa
     4021 gacgaaaggg cggcatgccg gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     4081 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc
     4141 ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     4201 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     4261 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     4321 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     4381 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     4441 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     4501 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     4561 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     4621 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     4681 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
     4741 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
     4801 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
     4861 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
     4921 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     4981 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     5041 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     5101 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     5161 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     5221 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     5281 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg
     5341 caccgggatc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     5401 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     5461 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga
     5521 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg
     5581 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg
     5641 ccgacgcgct gggctacgtc ttgctggcgt tcggctggca ttgaccctga gtgatttttc
     5701 tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg
     5761 catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta
     5821 cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg
     5881 cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg
     5941 agctggacgc ggatgaacag gcagacatct gtgaatcgct gcacgaccac gctgatgagc
     6001 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6061 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6121 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     6181 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6241 tagcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6301 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6361 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6421 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6481 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6541 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6601 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6661 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6721 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6781 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6841 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6901 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6961 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7021 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7081 tgagattatc aaaaaggatc ttcacctaga tccttttacg cgccctgtag cggcgcatta
     7141 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     7201 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     7261 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     7321 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     7381 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     7441 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     7501 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     7561 acgtttacaa tttaaatatt tgcttataca atcttcctgt ttttggggct tttctgatta
     7621 tcaaccgggg taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7681 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7741 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7801 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7861 ccggaagggc cgagcgcaga agtggtcctg caac
//
