LOCUS       sequence-341880-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S. Typhimurium gene stm2585 (sarA/steE) codon-optimized for 
            mammalian expression with N-terminal Flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  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 5993)
  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: pcDNA3.1-Flag-stm2585
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        163..542
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        543..746
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     697..717
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     791..810
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        791..809
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      863..872
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             872..895
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1512..1529)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1518..1742
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1788..2216
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1875..1894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2085..2106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2225..2245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2230..2559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2410..2545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2472..2491
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2626..3420
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2680..2699)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3290..3309
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3594..3715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3631..3650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3685..3704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3764..3780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3764..3780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3777..3799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3788..3804
                     /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(3812..3818,3819..3836,3837..3842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3857..3878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3995..4012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4166..4754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4246..4265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4925..5716,5717..5785))
                     /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     5548..5567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5786..5890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5965..5984)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
       61 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      121 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      181 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      241 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      301 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      361 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      421 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      481 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      541 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      601 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      661 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      721 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      781 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa
      841 gcttggtacc gagctcggat ccgccaccat ggactataag gacgacgacg ataaaatgat
      901 gcgatttgtc tacatctaca ttctggtcat ctatgggagc tacctgtggt tcagcctggg
      961 cgggaacatg ttcacaatca actcaactaa tagggtggcc agcaccattg ccccttacgc
     1021 ttgcgtgagc gacgtgaacc tggaagataa ggcaaccttc ctggacgagc acacatccat
     1081 ccatgccaat gatagctccc tgcagtgttt tgtgctgaac gaccagcacg tcccacagaa
     1141 tactctggct accgatgtgg aaggatacaa tcggggcctg caggagagaa tctccctgga
     1201 atatcagcca ctggagtcta tcgtgttcct gctggggact cccgctgtcc tggagaccaa
     1261 agaatcactg agcctgcccg tcagtcctga cgcactgaca cagaagctgc tgtctatttc
     1321 tagtaacgat gagtgcaaac tgtccggctc tactagttgt accacacccg ccagccataa
     1381 tcccccaagc ggctacattg ctcagtatcg gcacagcgca gaggtgtttc cagacgagta
     1441 agaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac
     1501 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1561 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1621 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1681 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1741 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1801 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1861 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1921 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1981 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2041 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2101 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2161 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2221 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2281 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2341 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2401 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2461 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2521 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2581 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2641 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2701 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2761 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2821 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2881 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2941 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3001 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3061 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3121 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3181 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3241 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3301 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3361 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3421 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3481 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3541 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3601 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3661 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3721 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3781 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3841 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4081 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4141 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4201 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4261 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4321 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4381 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4441 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4501 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4561 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4621 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4681 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4741 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4801 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4861 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4921 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4981 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5041 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5101 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5161 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5221 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5281 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5341 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5401 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5461 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5521 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5581 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5641 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5701 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5761 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5821 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5881 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     5941 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taa
//
