LOCUS       sequence-443904-        4559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes the active catalytic domain (amino acid 
            residues 164-468) of YopH. Inteded for co-expression with human Tyr 
            kinase plasmids to enhance bacterial kinase expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4559)
  AUTHORS   Albanese SK, Parton DL, Isik M, Rodriguez-Laureano L, Hanson SM, 
            Behr JM, Gradia S, Jeans C, Levinson NM, Seeliger MA, Chodera JD
  TITLE     An open library of human kinase domain constructs for automated 
            bacterial expression.
  JOURNAL   Biochemistry. 2018 Jul 13. doi: 10.1021/acs.biochem.7b01081.
  PUBMED    30004690
REFERENCE   2  (bases 1 to 4559)
  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: 
            "Biochemistry. 2018 Jul 13. doi: 10.1021/acs.biochem.7b01081."
COMMENT     Sequence Label: YopH (residues 164-468)
FEATURES             Location/Qualifiers
     source          1..4559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     558..577
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(597..674)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     720..739
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        720..738
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    743..766
                     /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             819..841
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1898..1916)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1912..1959
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2130..2921)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2922..3013)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(4073..4559,1..596))
                     /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"
ORIGIN
        1 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
       61 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
      121 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
      181 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
      241 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
      301 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
      361 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
      421 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
      481 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
      541 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
      601 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
      661 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaatt
      721 aatacgactc actataggga gagaattgtg agcggataac aattcccctg tagattaatt
      781 aagcggccgc cctgcaggac tcgagttcta gaaataattt tgtttaactt taagaaggag
      841 atatagatca tgcgtgaacg accacacact tctggccatc acggggctgg cgaagccaga
      901 gccaccgcac caagcactgt ttctccttat ggcccagaag cgcgcgcaga actcagcagc
      961 cgcctcacca cattgcgcaa tacgctggcg ccagcaacga atgatccgcg ttacttacaa
     1021 gcctgcggcg gtgaaaagct aaaccgattt agagatattc aatgctgtcg gcaaaccgca
     1081 gtacgcgccg atcttaatgc caattacatc caggtcggta acactcgtac catagcgtgc
     1141 cagtatccgc tacaatctca acttgaaagc catttccgta tgctggcaga aaaccgaacg
     1201 ccagtgttgg ctgttttagc gtccagttct gagatagcca atcaaagatt cggtatgcca
     1261 gattatttcc gccagagtgg tacctatggc agtatcactg tagagtctaa aatgactcag
     1321 caagttggtc tcggtgacgg gattatggca gatatgtata ctttaacgat tcgtgaagcg
     1381 ggtcaaaaaa caatctctgt tcctgtggtt catgttggca attggcccga tcagaccgca
     1441 gtcagctctg aagttaccaa ggcactcgct tcactggtag atcaaacagc agaaacaaaa
     1501 cgcaatatgt atgaaagcaa aggaagttca gcggtaggag atgactccaa attacggccg
     1561 gtaatacatt gccgtgcggg tgttggccgt actgcgcaac tgattggcgc aatgtgcatg
     1621 aatgatagtc gtaatagtca gttaagcgta gaagatatgg tcagccaaat gcgagtacaa
     1681 agaaatggta ttatggtaca aaaagatgag caacttgatg ttctgattaa gttggctgaa
     1741 ggacaagggc gaccattatt aaatagctaa taataagatc ccaactccat aaggatccgc
     1801 gatcgcggcg cgccacctgg tggccggccg gtaccacgcg tgcgcgctga tccggctgct
     1861 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1921 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag
     1981 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     2041 gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc ggatcttgcg
     2101 gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct
     2161 ttcacgtaat ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct
     2221 tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc
     2281 tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac
     2341 caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag
     2401 ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca
     2461 aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc
     2521 aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg
     2581 cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg
     2641 tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc
     2701 gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc
     2761 accgtaacca gcaaatcaat atcactgtgc ggcttcaggc cgccatccac tgcggagccg
     2821 tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct
     2881 gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcattat
     2941 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3001 aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag actgcggcgg
     3061 gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg
     3121 tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca
     3181 gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg
     3241 aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt
     3301 cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt
     3361 tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt
     3421 gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg
     3481 cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact
     3541 ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg
     3601 ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc
     3661 tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc
     3721 acctccccag gtggtttttt cgaaggttaa gtcagttggg gaactgctta accttcgatc
     3781 aaaccacctc cccaggtggt tttttcgttt acagggcaaa agattacgcg cagaaaaaaa
     3841 ggatctcaag aagatccttt gatcttttct actgaaccgc tcttatttca gtgcaattta
     3901 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
     3961 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     4021 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     4081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4141 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     4201 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     4261 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     4321 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     4381 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     4441 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     4501 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagcca
//
