LOCUS       sequence-257446-        6307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of ATP-grasp from Y. bercovieri.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6307)
  AUTHORS   Piasecka A, Czapinska H, Vielberg MT, Szczepanowski RH, Kiefersauer 
            R, Reed S, Groll M, Bochtler M
  TITLE     The Y. bercovieri Anbu crystal structure sheds light on the 
            evolution of highly (pseudo)symmetric multimers.
  JOURNAL   J Mol Biol. 2018 Mar 2;430(5):611-627. doi: 
            10.1016/j.jmb.2017.11.016. Epub 2017 Dec 16.
  PUBMED    29258816
REFERENCE   2  (bases 1 to 6307)
  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.jmb.2017.11.016"; journalName: "J Mol Biol"; date: 
            "2018-03-2- 2"; volume: "430"; issue: "5"; pages: "611-627"
COMMENT     Sequence Label: pQlinkH-YbATP-grasp
FEATURES             Location/Qualifiers
     source          1..6307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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)."
     protein_bind    53..69
                     /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             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             112..132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    145..169
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             169..189
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1633..1657)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1679..1773
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1867..2526
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1933..1952)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2591..2677
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2770..3852)
                     /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     3814..3833
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3853..3930)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(4014..4036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4122..4262
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4177..4196
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4277..4294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4448..5036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4528..4547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5207..5998,5999..6067))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5830..5849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6068..6172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6240..6258
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taactatgaa acatcaccat
      121 caccatcacc atggtgctgc aggtacaagt ttgtacaaaa aagcaggcga gaatctttat
      181 tttcagggat ccatgctgaa catcgacctt tccacatcac gatttttcga tgaaatgtta
      241 ctggcgcagg ggcaacaccg aagccattac gacgcctact ggcagtggtt acaacaggcc
      301 gatcaactga ccattgcgcg caagcgcgaa gaagctgcgc tgctgtttca ccgggtgggc
      361 attaccttca atgtgtatgg tgacgatgat ggcgccgagc gcttaatccc gtttgatagc
      421 gtgccacgta ttattcctgc cagtgaatgg cagatgttgg accgtggcat tcgccaacgt
      481 gttcaagcgc tgaatgcgtt tctgcatgat atttaccacg atcaacacat cctaaaagca
      541 gggattattc cggctgaaca agtgttagcc aatgaccaat atcagccctg tatgcaaggg
      601 gtgaacctgc accgcgacac ctacgcacat attattggcg tggatatggt tcgtaatagc
      661 gatggcctct attatgtgtt ggaggataat ctgcgcaccc catccggcgt ctcctatatg
      721 ctggaaaacc gcaaaatgat gatgcggctc tatccggagc tgtttgcgca acagcgcatc
      781 gcgccggttg agcgctaccc ttcccatttg ttgcaaacac tgcgcgaaag cacgccaatc
      841 aacgatccta ccgtggtggt gctgacccct gggcgcttta atagcgtcta ttttgagcac
      901 agttttttag cgcagcagat gggggtcgag ctagttgaaa gcgcggatct gttcgtcaaa
      961 gagggggcgg tctttatgcg caccaccgcc ggcccttgca agatagatgt tatctatcgg
     1021 cgagtggacg acgccttcct cgacccactg gctttccgcc cggactccat gctcggtgtc
     1081 cctgggctac tatcagtcta tcgggcaggt ggggtggtat tggctaacgc aattggtact
     1141 ggcgttgcag atgataaatc tatctatccg tatgtgccgg agatgatccg tttttacctc
     1201 gccgaagatc ctattctggc gaatgtcccc acctggcaat gccgccaacc tactgattta
     1261 tcttatgtct tggcgaatct ggatcagatg gtggtgaaag aagttcatgg cgcagggggc
     1321 tatggcatgt tgatcggccc cgcggccacc caagcggaga ttgcccaatt ccgccagttg
     1381 ctactggcgc gaccacaaaa ctacattgca caagagaccc tcgctctctc gacctgcccg
     1441 acctttgtaa acaacggcct tgccccacgc cacattgatc tgcgcccttt tgccctctcc
     1501 ggcgcggaaa ttcgtctagt gcccggtggc ctgacccgcg tggcactggc ggatggttca
     1561 ctggtggtga actcctcaca aggcggcggc accaaagaca cttgggtatt ggaggatgat
     1621 gcatgtccta ggacccagct ttcttgtaca aagtggtaag cttaattagc tgagcttgga
     1681 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     1741 ggttgccgcc gggcgttttt tattggtgag aatccaagta acaacaccat ttaaatggag
     1801 tggttacaaa tggagtggtt aattaaggct agcttggcga gattttcagg agctaaggaa
     1861 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1921 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1981 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     2041 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     2101 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     2161 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     2221 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     2281 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     2341 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     2401 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     2461 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     2521 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag
     2581 cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2641 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccctctagat tacgtgcagt
     2701 cgatgataag ctgtcaaaca tgagaattgt gcctaatgag tgagctaact tacattaatt
     2761 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     2881 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     2941 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     3001 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     3061 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     3121 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     3181 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     3241 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     3301 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     3361 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     3421 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     3481 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     3541 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     3601 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     3661 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     3721 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     3781 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     3841 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     3901 gcgaaaggtt ttgcaccatt cgatggtgtc ggaatttcgg gcagcgttgg gtcctggcca
     3961 cgggtgcgca tgatctagag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     4021 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4081 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     4141 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     4201 gagtgcacca catgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     4261 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4861 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6181 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6241 ataggcgtat cacgaggccc tttcgtcttc acctcgagct taattaacaa caccatttgt
     6301 cgagaaa
//
