LOCUS       sequence-372385-        7596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human STYK1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7596)
  AUTHORS   Salokas K, Liu X, Ohman T, Chowdhury I, Gawriyski L, Keskitalo S, 
            Varjosalo M
  TITLE     Physical and functional interactome atlas of human receptor tyrosine
            kinases.
  JOURNAL   EMBO Rep. 2022 Apr 5:e54041. doi: 10.15252/embr.202154041.
  PUBMED    35384245
REFERENCE   2  (bases 1 to 7596)
  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: "EMBO Rep. 
            2022 Apr 5:e54041. doi: 10.15252/embr.202154041."
COMMENT     Sequence Label: MAC_C_STYK1
FEATURES             Location/Qualifiers
     source          1..7596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        78..457
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        458..661
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     612..632
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    663..681
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    684..702
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     712..736
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    879..903
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2176..2200)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2247..2270
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2307..2330
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2361..2387
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2361..2387
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2388..2735,2736..2738,2739..3347)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(3391..3408)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3397..3621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3754..3773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3905..3952
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3960..4982
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3960..4982
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5112..5233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5149..5168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5203..5222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5282..5298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5282..5298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5295..5317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5306..5322
                     /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)."
     promoter        complement(join(5330..5336,5337..5354,5355..5360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5375..5396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5513..5530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5684..6272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5764..5783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6443..7234,7235..7303))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7066..7085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7304..7408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7483..7502)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
       61 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
      121 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      181 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      301 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      361 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      421 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      481 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      541 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      601 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      661 tctccctatc agtgatagag atctccctat cagtgataga gatcgtcgac gagctcgttt
      721 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      781 ccgggaccga tccagcctcc ggactctagc gtttaaactt aagcttggta ccgagctcgg
      841 atccactagt ccagtgtggt ggaattctgc agatatcaac aagtttgtac aaaaaagctg
      901 gcaccatggg catgacacgg atgctcctgg aatgcagtct cagtgacaag ttgtgtgtca
      961 tccaggagaa gcagtatgaa gtgattatcg tcccaacttt gttggttact atcttcctca
     1021 tccttcttgg ggtcatcctg tggcttttta tcagagaaca aagaactcaa cagcagcgtt
     1081 ctggacctca aggcattgcc cctgttcctc cacctaggga cctaagctgg gaagcaggac
     1141 atggaggaaa tgtggctttg ccacttaagg agacatccgt ggaaaacttt ctgggagcta
     1201 ccacacctgc cctggctaag ctgcaggtgc cgcgggagca actctctgaa gttctggagc
     1261 agatttgcag tggtagctgt gggcccatct ttcgagccaa tatgaacact ggggaccctt
     1321 ctaagcccaa gagtgttatt ctcaaggctt taaaagaacc agctgggctc catgaggtac
     1381 aagatttctt agggcgaatc caattccatc aatacctggg gaaacacaaa aacctggtgc
     1441 agctggaagg ctgctgcact gaaaagctgc cactctatat ggtgttggag gatgtggccc
     1501 agggggacct gctcggcttt ctctggacct gtcggcggga tgtgatgact atggatggtc
     1561 ttctctatga tctcacagaa aaacaagtat atcacatcgg aaagcaggtc cttttggcgc
     1621 tggaattcct gcaggagaag catttgttcc atggggatgt ggcagccagg aatattctga
     1681 tgcaaagtga tctcactgct aagctctgtg gattaggcct ggcttatgaa gtttacaccc
     1741 gaggggccat ctcctctact caaaccatac ctctcaagtg gcttgcccca gaacggcttc
     1801 tcctgagacc tgctagcatc agagcagatg tctggtcttt tgggatcctg ctctatgaga
     1861 tggtgactct aggagcacca ccgtatcctg aagtccctcc taccagcatc ctagagcatc
     1921 tccaaagaag gaaaatcatg aagagaccca gtagctgcac acataccatg tacagtatca
     1981 tgaagtcctg ctggcgctgg cgtgaggctg accgcccctc acctagagag ctgcgcttgc
     2041 gcctagaagc tgccattaaa actgcagatg acgaggctgt gttacaagta ccagagttgg
     2101 tggtacctga actgtatgca gctgtggccg gcatcagagt ggagagcctc ttctacaact
     2161 atagcatgct tggcgaccca gctttcttgt acaaagtggt tgatatcaag gccgccgaca
     2221 tcaccagcct gtacaagaaa gtggggtgga gccatcccca gttcgagaag ggcggaggat
     2281 ctggcggcgg aagcggcgga ggaagctggt cccaccctca gtttgagaaa ggcaccgagc
     2341 tgggcagcac catggccagc tacccctacg acgtgcccga ctacgccaag gacaacaccg
     2401 tgcccctgaa gctgatcgcc ctgctggcca acggcgagtt ccactctggc gagcagctgg
     2461 gcgagacact gggcatgagc agagccgcca tcaacaagca catccagacc ctgcgggact
     2521 ggggcgtgga cgtgttcaca gtgcccggca agggctacag cctgcccgag cccatccagc
     2581 tgctgaacgc caagcagatc ctgggccagc tggatggcgg atctgtggcc gtgctgcccg
     2641 tgatcgacag caccaaccag tatctgctcg accggatcgg cgagctgaag tccggcgacg
     2701 cctgtatcgc cgagtaccag caggccggca gaggcggcag gggcagaaag tggttcagcc
     2761 ccttcggcgc caacctgtac ctgagcatgt tctggagact ggaacagggc ccagccgccg
     2821 ctattggcct gagcctggtc atcggcatcg tgatggccga ggtgctgcgg aagctgggcg
     2881 ccgacaaagt gcgcgtgaag tggcccaacg acctgtatct gcaggaccgg aagctggccg
     2941 gcatcctggt ggaactgacc ggcaaaacag gcgacgccgc ccagatcgtg atcggcgctg
     3001 gcatcaacat ggccatgcgg cgggtggaag agagcgtggt caaccagggc tggatcaccc
     3061 tgcaggaagc cggcatcaac ctggaccgga acaccctggc cgccatgctg atcagagagc
     3121 tgagagccgc cctggaactg ttcgagcagg aaggcctggc cccctacctg agcagatggg
     3181 agaagctgga caacttcatc aaccggcccg tgaaactgat catcggcgac aaagagatct
     3241 tcggcatcag ccggggcatc gacaagcagg gggccctgct cctggaacag gacggcatca
     3301 tcaagccctg gatgggcggc gagatcagcc tgcggagcgc cgagaagtga tgactcgagt
     3361 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3421 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3481 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3541 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3601 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3661 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3721 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3781 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3841 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3901 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
     3961 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     4021 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     4081 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     4141 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     4201 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     4261 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     4321 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     4381 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     4441 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     4501 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     4561 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     4621 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     4681 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     4741 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     4801 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     4861 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     4921 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     4981 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5041 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5101 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5161 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5221 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5281 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5341 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5401 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5461 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5521 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5581 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5641 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6121 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6721 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7261 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7321 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7381 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7441 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     7501 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7561 gagcaaaatt taagctacaa caaggcaagg cttgac
//
