LOCUS       sequence-341890-        8237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of sarA with its GSK3-binding SxxxSP motif 
            mutated to SxxxAP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8237)
  AUTHORS   Gibbs KD, Washington EJ, Jaslow SL, Bourgeois JS, Foster MW, Guo R, 
            Brennan RG, Ko DC
  TITLE     The Salmonella Secreted Effector SarA/SteE Mimics Cytokine Receptor 
            Signaling to Activate STAT3.
  JOURNAL   Cell Host Microbe. 2020 Jan 8;27(1):129-139.e4. doi: 
            10.1016/j.chom.2019.11.012. Epub 2019 Dec 31.
  PUBMED    31901521
REFERENCE   2  (bases 1 to 8237)
  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: 
            "10.1016/j.chom.2019.11.012"; journalName: "Cell Host Microbe"; 
            date: "2020-01-8- 8"; volume: "27"; issue: "1"; pages: "129-139.e4"
COMMENT     Sequence Label: pWSK129-stm2585_S130A
FEATURES             Location/Qualifiers
     source          1..8237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(460..479)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             471..1286
                     /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             471..1286
                     /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(544..563)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2696..2713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2830..2851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2866..2871,2872..2889,2890..2896)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2904..2920
                     /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     2909..2931
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2928..2944
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2928..2944
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2963..2983
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2965..2983
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4398..4417)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4399..4417)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4427..4444)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4427..4443)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4436..4458)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4585..5013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4672..4691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4882..4903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6013..6235
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             6283..7233
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 gaggacaagg ctacattttt agatgaacat acctcaatac atgcgaatga ttcttctttg
       61 caatgctttg tcttaaatga tcagcatgtt ccgcagaata cattagctac agatgttgaa
      121 ggctacaata gaggattaca ggaacgtata agcctcgagt accagcccct ggaaagcatt
      181 gtttttctac tcggcacccc tgcagtttta gagactaaag agtctttatc attaccagtt
      241 gccccggatg ctttaaccca aaaactatta agcattagta gcaatgatga atgtaaatta
      301 tcaggtagca ccagttgcac tacccatcct tcaactcagc aaaagttcga tttattcaac
      361 aaagccacgt tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
      421 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
      481 ttcaacggga aacgtcttgc tcgaggccgc gattaaattc caacatggat gctgatttat
      541 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
      601 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
      661 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
      721 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggga
      781 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
      841 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
      901 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
      961 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1021 agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1081 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1141 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     1201 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     1261 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     1321 gcattacgct gacttgacgg gacggcggct ttgttgaata aatcgaactt ttgctgagtt
     1381 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt
     1441 caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt
     1501 ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg
     1561 cagacctcag cgctattctg accttgccat cacgactgtg ctggtcatta aacgcgtatt
     1621 caggctgacc ctgcgcgctg cgcagggctt tattgattcc atttttacac tgatgaatgt
     1681 tccgttgcgc tgcccggatt acctgcatat taaagagctg tatgtttctc agtgcgatcg
     1741 aaataagggc acgggtaaag cgataatgcg ctttatagcc cggcttgcgc ttgaacagga
     1801 atgccttagc cttagctgga acgctgaaaa atccaacccc ggcgctaacc gtttttatca
     1861 ggctctggga ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct
     1921 gagcaaactg gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     1981 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     2041 ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc
     2101 gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg
     2161 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     2221 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     2281 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     2341 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     2401 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     2461 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     2521 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     2581 catacggaat tgatctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2641 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2701 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2761 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2821 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     2881 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     2941 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     3001 ccaccgcggt ggcggccgct ctagactaca tgccaacttt acttagtatc atattttctt
     3061 gctacatatc tgtttggcaa gggtatgaat aaaaaacaag attaatttaa ttgatatcgc
     3121 aacagataag cacctgcatt tatctttaaa cttttttatg gccaaccaga tttttcacca
     3181 tctgcgcacg tttattcggg catgcgttga aaacctgcaa aaaaagaaaa atatcgatgc
     3241 ttattatttt ttctttaatt aaattttcgc tcaacaaact taattattaa tccaatgaca
     3301 gtgaagtgtg aactatgctg agcataaacg acatcaatag caagggcaag ggcaagggca
     3361 agggcaaggg caagggcaag ggcaagggca agggcaatct gagtatttac aggtgaaatt
     3421 atgaaaaata gtatattttt accattgctt ctggtgctgt cggcaacgac cttttcggca
     3481 ttggtgatgg ctgccagtca ttctccccac cagataatac aaaacatttt tccagtggct
     3541 ggccgccaga tatcctgaag caactgaatt ggtctcaggt ttgtggttgt tagtatactg
     3601 aatattactt caggatggca tgttttattg acaccagcct gatttttcac aacgttatat
     3661 tacccgttta cagataattc ccagctaata aatcaggaag ttccagatgt aacagaagtt
     3721 atgaggtgta ttaattgata ggtctaacag gcttgaatta cttttcttta aaaactactg
     3781 catgtaaaag ggtctcctct tgttgtgatg agattcgtat atatttatat cttagtgatt
     3841 tatggatcat acttgtggtt ttccttagga ggtaacatgt ttacaattaa tagtactaac
     3901 agggtggcga gcacaatcgc tccatatgca tgtgtaagcg atgttaattt agaggacaag
     3961 gctacatttt tagatgaaca tacctcaata catgcgaatg attcttcttt gcaatgcttt
     4021 gtcttaaatg atcagcatgt tccgcagaat acattagcta cagatgttga aggctacaat
     4081 agaggattac aggaacgtat aagcctcgag taccagcccc tggaaagcat tgtttttcta
     4141 ctcggcaccc ctgcagtttt agagactaaa gagtctttat cattaccagt tgccccggat
     4201 gctttaaccc aaaaactatt aagcattagt agcaatgatg aatgtaaatt atcaggtagc
     4261 accagttgca ctaccccagc cagtcataac ccaccgtccg gttatatcgc tcaatacagg
     4321 cattctgcag aggttttccc ggatgaataa aacaaagcgt tataaatagc tgcgccgagg
     4381 gcccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     4441 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4501 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4561 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4621 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4681 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4741 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4801 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4861 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4921 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4981 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aattcgaccg atgcccttga
     5041 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     5101 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     5161 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     5221 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     5281 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
     5341 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
     5401 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
     5461 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga ccgctgatcg
     5521 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg
     5581 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
     5641 cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccgc agacccgcca
     5701 taaaacgccc tgagaagccc gtgacgggct tttcttgtat tatgggtagt ttccttgcat
     5761 gaatccataa aaggcgcctg tagtgccatt tacccccatt cactgccaga gccgtgagcg
     5821 cagcgaactg aatgtcacga aaaagacagc gactcaggtg cctgatggtc ggagacaaaa
     5881 ggaatattca gcgatttgcc cgagcttgcg agggtgctac ttaagccttt agggttttaa
     5941 ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact
     6001 gactaaagta gtgagttata cacagggctg ggatctattc tttttatctt tttttattct
     6061 ttctttattc tataaattat aaccacttga atataaacaa aaaaaacaca caaaggtcta
     6121 gcggaattta cagagggtct agcagaattt acaagttttc cagcaaaggt ctagcagaat
     6181 ttacagatac ccacaactca aaggaaaagg actagtaatt atcattgact agcccatctc
     6241 aattggtata gtgattaaaa tcacctagac caattgagat gtatgtctga attagttgtt
     6301 ttcaaagcaa atgaactagc gattagtcgc tatgacttaa cggagcatga aaccaagcta
     6361 attttatgct gtgtggcact actcaacccc acgattgaaa accctacaag gaaagaacgg
     6421 acggtatcgt tcacttataa ccaatacgct cagatgatga acatcagtag ggaaaatgct
     6481 tatggtgtat tagctaaagc aaccagagag ctgatgacga gaactgtgga aatcaggaat
     6541 cctttggtta aaggctttga gattttccag tggacaaact atgccaagtt ctcaagcgaa
     6601 aaattagaat tagtttttag tgaagagata ttgccttatc ttttccagtt aaaaaaattc
     6661 ataaaatata atctggaaca tgttaagtct tttgaaaaca aatactctat gaggatttat
     6721 gagtggttat taaaagaact aacacaaaag aaaactcaca aggcaaatat agagattagc
     6781 cttgatgaat ttaagttcat gttaatgctt gaaaataact accatgagtt taaaaggctt
     6841 aaccaatggg ttttgaaacc aataagtaaa gatttaaaca cttacagcaa tatgaaattg
     6901 gtggttgata agcgaggccg cccgactgat acgttgattt tccaagttga actagataga
     6961 caaatggatc tcgtaaccga acttgagaac aaccagataa aaatgaatgg tgacaaaata
     7021 ccaacaacca ttacatcaga ttcctaccta cataacggac taagaaaaac actacacgat
     7081 gctttaactg caaaaattca gctcaccagt tttgaggcaa aatttttgag tgacatgcaa
     7141 agtaagtatg atctcaatgg ttcgttctca tggctcacgc aaaaacaacg aaccacacta
     7201 gagaacatac tggctaaata cggaaggatc tgaggttctt atggctcttg tatctatcag
     7261 tgaagcatca agactaacaa acaaaagtag aacaactgtt caccgttaca tatcaaaggg
     7321 aaaactgtcc atatgcacag atgaaaacgg tgtaaaaaag atagatacat cagagctttt
     7381 acgagttttt ggtgcattca aagctgttca ccatgaacag atcgacaatg taacagatga
     7441 acagcatgta acacctaata gaacaggtga aaccagtaaa acaaagcaac tagaacatga
     7501 aattgaacac ctgagacaac ttgttacagc tcaacagtca cacatagaca gcctgaaaca
     7561 ggcgatgctg cttatcgaat caaagctgcc gacaacacgg gagccagtga cgcctcccgt
     7621 ggggaaaaaa tcatggcaat tctggaagaa atagcgcttt cagccggcaa accggctgaa
     7681 gccggatctg cgattctgat aacaaactag caacaccaga acagcccgtt tgcgggcagc
     7741 aaaacccgta cttttggacg ttccggcggt tttttgtggc gagtggtgtt cgggcggtgc
     7801 gcgcaagatc cattatgtta aacgggcgag tttacatctc aaaaccgccc gcttaacacc
     7861 atcagaaatc ctcagcgcga ttttaagcac caaccccccc ccgtaacacc caaatccata
     7921 ctgaaagtgg ctttgttgaa taaatcagat ttcgggtaag tctcccccgt agcgggttgt
     7981 gttttcaggc aatacgcacg ctttcaggca tacctgcttt cgtcattttg ttcagcgctc
     8041 gtaccagggc catagcctcc gcaacctgac catcgtagtc acgcagcgtc agtgaacccc
     8101 cgaacagctg taatccgggc agcgcaacgg aacattcatc agtgtaaaaa tggaatcaat
     8161 aaagccctgc gcagcgcgca gggctttatt gattccattt ttacactgat gaatgttccg
     8221 ttgcgctgcc cggatta
//
