LOCUS       sequence-215709-        5664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express murine Syk (N)SH2 domain (Ser 8 to Gly 117), with an 
            N-terminal (His)6 tag and TEV cleavage site, in E.coli strain 
            Rosetta 2 (DE3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  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 5664)
  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(+)-N-SH2
FEATURES             Location/Qualifiers
     source          1..5664
                     /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"
     CDS             complement(552..572)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(573..590)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             601..623
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    638..662
                     /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(662..681)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(663..681)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(769..788)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        994..1071
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1072..2154
                     /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(1091..1110)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2963..3154
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3148..3170)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3256..3395
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3311..3330
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3410..3427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3581..4169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3661..3680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4280..4299)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4291..5106
                     /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             4291..5106
                     /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(4364..4383)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5198..5653)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5335..5356)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5547..5566
                     /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 gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgatatat atgagctctt atccggtctt gggctgtact
      241 cccgggggcc ggttgaaggg cttcttaagg aggcagataa ggccatcagg ttcctgggag
      301 tggtaatggc agaggtctgc tgggctggca tgggccctgc ccccggagat ggcgtaggtg
      361 ccattaagtt ccctctcgat ggtgtagtgg tgtgccttcc tgttgtgagc caccgacaaa
      421 gcaaaaccac ccaggtaatt gcggctctgg cgtagcaggt agagcccatc ggtcatgcct
      481 ccctggacca ggtagtcttc agcctcttcc cgggtgatgt tgccaaaaaa gtaggtcagg
      541 tggttggcgc tgccctgaaa gtacaggttt tcgtggtggt ggtggtggtg catatgtata
      601 tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt
      661 cccctatagt gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc ctctacgccg
      721 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
      781 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
      841 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
      901 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
      961 aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt
     1021 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     1081 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     1141 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     1201 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     1261 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     1321 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     1381 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     1441 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     1501 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     1561 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     1621 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     1681 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     1741 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     1801 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     1861 gttggtgcgg acatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     1921 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     1981 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     2041 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2101 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2161 cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag
     2221 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2281 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
     2341 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
     2401 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
     2461 cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct
     2521 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc
     2581 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac
     2641 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg
     2701 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
     2761 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat
     2821 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt
     2881 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa
     2941 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg
     3001 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga
     3061 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct
     3121 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     3181 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     3241 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
     3301 cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg tgtgaaatac
     3361 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4021 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat aaaactgtct
     4261 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     4321 tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     4381 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     4441 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     4501 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     4561 cctgatgatg catggttact caccactgcg atccccggga aaacagcatt ccaggtatta
     4621 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     4681 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     4741 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     4801 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     4861 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     4921 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     4981 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     5041 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     5101 ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5161 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     5221 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5281 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5341 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5401 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5461 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5521 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5581 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5641 ctacagggcg cgtcccattc gcca
//
