LOCUS       sequence-342057-        4301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intended for co-expression with human Ser/Thr kinase plasmids to 
            enhance bacterial kinase expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4301)
  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 4301)
  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: Lambda Phosphatase
FEATURES             Location/Qualifiers
     source          1..4301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(583..1665)
                     /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     1627..1646
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1666..1743)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1789..1808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1789..1807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1812..1835
                     /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             1888..1910
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2709..2727)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2723..2770
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2941..3732)
                     /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(3733..3824)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact ggcagcagcc
       61 attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg ctcttgaagt
      121 gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc tgcgaaagcc
      181 agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc acctccccag
      241 gtggtttttt cgaaggttaa gtcagttggg gaactgctta accttcgatc aaaccacctc
      301 cccaggtggt tttttcgttt acagggcaaa agattacgcg cagaaaaaaa ggatctcaag
      361 aagatccttt gatcttttct actgaaccgc tcttatttca gtgcaattta tctcttcaaa
      421 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
      481 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
      541 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
      601 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      661 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
      721 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
      781 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
      841 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
      901 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
      961 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     1021 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     1081 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     1141 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     1201 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     1261 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     1321 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     1381 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     1441 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     1501 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     1561 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     1621 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     1681 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     1741 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca
     1801 ctatagggag agaattgtga gcggataaca attcccctgt agattaatta agcggccgcc
     1861 ctgcaggact cgagttctag aaataatttt gtttaacttt aagaaggaga tatagatcat
     1921 gcggtactac gaaaaaatcg acggtagcaa atatcgcaat atctgggttg tgggcgacct
     1981 gcatggctgc tatacaaatc tgatgaacaa gcttgacact atcgggttcg ataataaaaa
     2041 agacctgctg atcagcgtgg gagatctggt tgaccgtggg gctgaaaacg ttgaatgcct
     2101 ggaactgatc accttcccct ggttccgcgc cgtgcgtggc aaccatgaac agatgatgat
     2161 cgatggactg agcgaacgcg gtaatgtgaa ccattggctg ttaaatggag gaggctggtt
     2221 tttcaatctg gactacgaca aagagatcct ggccaaagcc ttggcacata aagcagatga
     2281 gctgcctctc atcattgaac tggttagtaa agataaaaaa tatgtgattt gccacgcgga
     2341 ttaccctttt gacgagtatg aatttggaaa accggtggat catcagcagg ttatttggaa
     2401 ccgcgagcgc atctctaatt cgcagaatgg catcgtgaag gaaattaagg gcgccgatac
     2461 cttcatcttc ggtcatacgc cagcagtgaa acccctgaag tttgcgaacc agatgtatat
     2521 tgataccggc gcggttttct gcggtaattt gacgctcatc caggttcagg gcgaaggtgc
     2581 ctaataagat cccaactcca taaggatccg cgatcgcggc gcgccacctg gtggccggcc
     2641 ggtaccacgc gtgcgcgctg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     2701 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     2761 gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     2821 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     2881 gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca
     2941 ttatttgccg actaccttgg tgatctcgcc tttcacgtaa tggacaaatt cttccaactg
     3001 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
     3061 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     3121 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     3181 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     3241 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     3301 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
     3361 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     3421 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     3481 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     3541 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     3601 cggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
     3661 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
     3721 cgcttccctc atactcttcc tttttcatta ttattgaagc atttatcagg gttattgtct
     3781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta gctcactcgg
     3841 tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac
     3901 agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg
     3961 gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt
     4021 gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag
     4081 gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac
     4141 cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct
     4201 catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta
     4261 agcaagaact ccccgttcag cccgactgct gcgccttatc c
//
