LOCUS       sequence-215610-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express murine Syk tandem SH2 domains (Ser 8 to Gln 264), with Y130E
            mutation to mimic phosphorylation, in E.coli strain Rosetta 2 (DE3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  AUTHORS   Feng C, Post CB
  TITLE     Insights into the allosteric regulation of Syk association with 
            receptor ITAM, a multi-state equilibrium.
  JOURNAL   Phys Chem Chem Phys. 2016 Feb 17;18(8):5807-18. doi: 
            10.1039/c5cp05417f.
  PUBMED    26468009
REFERENCE   2  (bases 1 to 6011)
  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"; journalName: 
            "Phys Chem Chem Phys"; date: "2016-02-17- 17"; volume: "18"; issue: 
            "8"; pages: "5807-18"
COMMENT     Sequence Label: pET-30a(+)-Syk-tSH2-PM
FEATURES             Location/Qualifiers
     source          1..6011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             948..970
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    985..1009
                     /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(1009..1028)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1010..1028)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1116..1135)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1341..1418
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1419..2501
                     /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(1438..1457)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3310..3501
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3495..3517)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3603..3742
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3658..3677
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3757..3774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3928..4516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4008..4027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4627..4646)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4638..5453
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4638..5453
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4711..4730)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5545..6000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5682..5703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5894..5913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttattgt gcaccgatct
      181 tctggcatgg taccgtgagg actcttagta gcccatctgg cttgtaagag taatgttcca
      241 ctagctgcca gagggtgtca aacttcttcc cctcaggaat ggagagcttc ccggtcttgt
      301 ccctgtcaat gcggtagtgc aatactttcc cttcgtgcag caggcacaga gcataggagc
      361 cgctgttgtc tctggccctg atcaggaatt ttccattggt ctttgaccct atgaggaccg
      421 tctgctctga ttcatctctg gagatgttgc catggaacca gggcatcttc tcatgggccg
      481 tggtggcgat cagcttctcc agctggggct tctggctgat gatggcttgc tccagagcct
      541 ggccctgaag gttccaggtc tgtttcacct cttccctgat gaggttctcc ttcaggtcct
      601 caaagggtcc ggtcttgggc tgtactcccg ggggccggtt gaagggcttc ttaaggaggc
      661 agataaggcc atcaggttcc tgggagtggt aatggcagag gtctgctggg ctggcatggg
      721 ccctgccccc ggagatggcg taggtgccat taagttccct ctcgatggtg tagtggtgtg
      781 ccttcctgtt gtgagccacc gacaaagcaa aaccacccag gtaattgcgg ctctggcgta
      841 gcaggtagag cccatcggtc atgcctccct ggaccaggta gtcttcagcc tcttcccggg
      901 tgatgttgcc aaaaaagtag gtcaggtggt tggcgctcat atgtatatct ccttcttaaa
      961 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag
     1021 tcgtattaat ttcgcgggat cgagatcgat ctcgatcctc tacgccggac gcatcgtggc
     1081 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     1141 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
     1201 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
     1261 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
     1321 gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
     1381 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     1441 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     1501 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     1561 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     1621 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     1681 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     1741 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     1801 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     1861 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     1921 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     1981 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     2041 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     2101 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     2161 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggaca
     2221 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     2281 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     2341 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     2401 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2461 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
     2521 agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca
     2581 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     2641 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     2701 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     2761 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     2821 attatcgccg gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
     2881 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
     2941 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
     3001 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
     3061 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     3121 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
     3181 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
     3241 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
     3301 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
     3361 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
     3421 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
     3481 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3541 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3601 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     3661 catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt
     3721 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3781 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4561 cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct tacataaaca
     4621 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct aggccgcgat
     4681 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     4741 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     4801 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     4861 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     4921 ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg
     4981 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     5041 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     5101 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     5161 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     5221 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     5281 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     5341 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     5401 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taagaattaa
     5461 ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5521 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     5581 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5641 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5701 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5761 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5821 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5881 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5941 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6001 cccattcgcc a
//
