LOCUS       sequence-341887-        5286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Native S. Typhimurium gene stm2585 (sarA/steE) with N-terminal GFP 
            and Flag tags and its GSK3-binding SxxxSP motif mutated to SxxxAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5286)
  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 5286)
  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: pEGFP-Flag-stm2585_S130A
FEATURES             Location/Qualifiers
     source          1..5286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..172
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      230..239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             239..262
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    959..1080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(996..1015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1050..1069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1087..1542)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1224..1245)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1436..1455
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1569..1673
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1675..2032
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1675..1866
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1698..1718)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1883..2018
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2067..2861
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2067..2861
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2121..2140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2731..2750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3049..3068)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3093..3140
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3469..4057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3958..3977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4232..4535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4536..4739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4690..4710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4778..4787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4784..4786,4787..4789,4790..5286,1..214)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4784..4786,4787..4789,4790..5286,1..214)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4829..4850)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5090..5109)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
       61 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
      121 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      181 cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatccg ccaccatgga
      241 ctacaaagac gatgacgaca agatgatgag attcgtatat atttatatct tagtgattta
      301 tggatcatac ttgtggtttt ccttaggagg taacatgttt acaattaata gtactaacag
      361 ggtggcgagc acaatcgctc catatgcatg tgtaagcgat gttaatttag aggacaaggc
      421 tacattttta gatgaacata cctcaataca tgcgaatgat tcttctttgc aatgctttgt
      481 cttaaatgat cagcatgttc cgcagaatac attagctaca gatgttgaag gctacaatag
      541 aggattacag gaacgtataa gcctcgagta ccagcccctg gaaagcattg tttttctact
      601 cggcacccct gcagttttag agactaaaga gtctttatca ttaccagttg ccccggatgc
      661 tttaacccaa aaactattaa gcattagtag caatgatgaa tgtaaattat caggtagcac
      721 cagttgcact accccagcca gtcataaccc accgtccggt tatatcgctc aatacaggca
      781 ttctgcagag gttttcccgg atgaataagt cgacggtacc gcgggcccgg gatccaccgg
      841 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
      901 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
      961 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1021 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1081 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1141 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1201 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1261 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1321 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1381 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     1441 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     1501 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     1561 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1621 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     1681 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1741 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1801 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1861 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1921 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     1981 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     2041 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2101 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2161 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2221 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     2281 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2341 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2401 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2461 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     2521 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     2581 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     2641 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2701 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     2761 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     2821 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     2881 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     2941 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3001 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     3061 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3121 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3181 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3241 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3301 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3361 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3421 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4081 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4141 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     4201 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4261 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4321 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4381 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4441 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4501 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     4561 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4621 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4681 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     4741 gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga
     4801 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4861 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     4921 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     4981 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     5041 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     5101 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     5161 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     5221 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     5281 caagat
//
