LOCUS       sequence-459259-        7143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human SYK Kinase in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7143)
  TITLE     Mesecar lab plasmids 2024
REFERENCE   2  (bases 1 to 7143)
  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"
COMMENT     Sequence Label: pET28a_SYK
FEATURES             Location/Qualifiers
     source          1..7143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             14..31
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             41..61
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1922..1939
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1992..2010)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2006..2053
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2090..2545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2177..2196)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2387..2408
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2637..3452)
                     /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             complement(2637..3452)
                     /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     3360..3379
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3444..3463
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3574..4162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4063..4082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4316..4333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4348..4487
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4413..4432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4573..4595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4589..4780)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5589..6671)
                     /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     6633..6652
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6672..6749)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6955..6974
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7058..7077
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7058..7076
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7077..7101
                     /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             7116..7138
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 catgggcagc agccatcatc atcatcatca cagcagcggc gagaatcttt attttcaggg
       61 ccatatgatg gcgtcaagtg gaatggctga ttctgcaaac cacctgccgt tctttttcgg
      121 taacattacc cgtgaagagg cggaagacta tttggtccag ggcggtatga gcgatggtct
      181 ctacctgtta cgtcaaagcc gtaattatct gggtggtttt gcgctgtccg tggcccacgg
      241 ccgtaaagcg caccactaca ccattgaacg tgagctgaat ggcacctatg cgatcgccgg
      301 tggtcggacc catgcatccc cggctgacct gtgccattac catagccaag agtccgacgg
      361 cttggtatgt ttgctgaaaa agccgttcaa ccgtccgcaa ggcgttcagc caaaaactgg
      421 tccgtttgaa gacctgaaag aaaatctgat ccgtgagtac gtgaaacaaa cctggaatct
      481 gcagggtcag gcgttggagc aagctatcat cagccagaaa ccgcagttgg aaaagctgat
      541 cgcgaccacc gctcacgaga agatgccgtg gtttcatggt aagatctctc gcgaggaatc
      601 ggaacaaatc gtgctgattg gtagtaaaac gaacggcaaa ttcctgattc gcgcacgtga
      661 taacaacggc tcttatgcct tatgcctgct tcacgagggc aaagttctgc actaccgcat
      721 tgataaagat aagacgggta agctgtctat tccggagggc aagaagttcg ataccctgtg
      781 gcagctagtg gaacattaca gctacaaggc ggacggtttg ctacgcgtgc tgaccgttcc
      841 gtgccagaaa atcggcacgc agggcaacgt taatttcggc ggtcgtccgc aattgccagg
      901 ttcacacccg gctagcagcc cggcgcaggg caaccgccag gagagcaccg ttagctttaa
      961 cccgtatgaa ccggaactcg cgccttgggc agcggacaag ggtccgcaac gtgaggcgct
     1021 gccgcatatg ctcgagatgg atacagaagt atacgagtca ccctatgctg acccggaaga
     1081 gattcgtcca aaagaggtgt atctggaccg taaactgttg accctggaag ataaggagct
     1141 gggctcgggc aacttcggca ctgttaaaaa aggctattac caaatgaaaa aggttgttaa
     1201 gaccgtcgcc gttaagatcc tgaaaaacga agccaatgat ccggcactga aagatgagtt
     1261 gctggcggag gcgaacgtaa tgcagcagct ggacaacccg tatattgttc gtatgattgg
     1321 tatctgtgag gctgagagct ggatgctggt gatggagatg gcagagttgg gccctctcaa
     1381 taaatacttg caacaaaacc gccacgtgaa ggacaagaat atcatcgaac tggtgcacca
     1441 ggtgtcaatg ggtatgaaat acttggaaga atctaatttc gtgcaccgtg atctggctgc
     1501 gcgtaacgtt ctgcttgtta cccagcatta tgcgaaaatc tccgattttg gtctgagcaa
     1561 ggccttgcgc gcggatgaaa attattacaa ggcgcagacc catggcaagt ggccggttaa
     1621 gtggtatgca ccggaatgta ttaactatta caagtttagc agcaaatccg acgtgtggtc
     1681 tttcggagtc ctgatgtggg aagcgttcag ctacggccaa aagccgtacc gcggtatgaa
     1741 aggtagcgag gtcacggcta tgctagagaa aggtgaacgt atgggctgcc cggcgggttg
     1801 cccgagagaa atgtatgatc tgatgaacct ttgctggacc tacgacgtgg agaatcgacc
     1861 gggttttgcg gcagttgagc tccgcctgcg taactactac tacgacgtgg tgaacctcga
     1921 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     1981 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     2041 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     2101 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2161 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2221 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2281 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2341 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2401 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2461 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2521 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2581 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
     2641 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     2701 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     2761 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     2821 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     2881 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     2941 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     3001 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     3061 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     3121 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     3181 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     3241 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     3301 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     3361 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     3421 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     3481 gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3721 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4201 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4261 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4321 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4381 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4441 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     4501 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4561 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4621 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     4681 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4741 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     4801 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4861 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4921 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     4981 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     5041 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     5101 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     5161 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     5221 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
     5281 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     5341 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     5401 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     5461 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     5521 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     5581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     5701 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     5761 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     5821 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     5881 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     5941 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     6001 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     6061 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     6121 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     6181 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     6241 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     6301 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     6361 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     6421 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     6481 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     6541 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     6601 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     6661 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     6721 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     6781 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     6841 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     6901 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     6961 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     7021 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     7081 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     7141 tac
//
