LOCUS       sequence-360223-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminally tagged Salmonella Typhimurium SopB for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  AUTHORS   Walpole GFW, Pacheco J, Chauhan N, Clark J, Anderson KE, Abbas YM, 
            Brabant-Kirwan D, Montano-Rendon F, Liu Z, Zhu H, Brumell JH, 
            Deiters A, Stephens LR, Hawkins PT, Hammond GRV, Grinstein S, Fairn 
            GD
  TITLE     Kinase-independent synthesis of 3-phosphorylated phosphoinositides 
            by a phosphotransferase.
  JOURNAL   Nat Cell Biol. 2022 Apr 28. pii: 10.1038/s41556-022-00895-y. doi: 
            10.1038/s41556-022-00895-y.
  PUBMED    35484249
REFERENCE   2  (bases 1 to 6401)
  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: "Nat Cell 
            Biol. 2022 Apr 28. pii: 10.1038/s41556-022-00895-y. doi: 
            10.1038/s41556-022-00895-y."
COMMENT     Sequence Label: SopB-EGFP
FEATURES             Location/Qualifiers
     source          1..6401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..377
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        378..581
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     532..552
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2354..2363
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2360..2362,2363..2365,2366..3079)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2360..2362,2363..2365,2366..3076)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2405..2426)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2666..2685)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3013..3034
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3202..3323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3239..3258)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3293..3312
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3330..3785)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3467..3488)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3679..3698
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3812..3916
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3918..4275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3918..4109
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3941..3961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4126..4261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4310..5104
                     /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             4310..5104
                     /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(4364..4383)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4974..4993
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5292..5311)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5336..5383
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5712..6300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6201..6220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      421 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      481 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      541 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      601 tccgctagcg ctaccggact cagatctcga gctcaagctt cgaattctaa tgcaaataca
      661 gagcttctat cactcagctt cactaaaaac ccaggaggct tttaaaagcc tacaaaaaac
      721 cttatacaac ggaatgcaga ttctctcagg ccagggcaaa gcgccggcta aagcgcccga
      781 cgctcgcccg gaaattattg tcctgcgaga acccggcgcg acatggggga attatctaca
      841 gcatcagaag gcgtctaacc actcgctgca taacctctat aacttacagc gcgatcttct
      901 taccgtcgcg gcaaccgttc tgggtaaaca agacccggtt ctaacgtcaa tggcaaacca
      961 aatggagtta gccaaagtta aagcggaccg gccagcaaca aaacaagaag aagccgcggc
     1021 aaaagcattg aagaaaaatc ttatcgaact tattgcagca cgcactcagc agcaggatgg
     1081 cttacctgca aaagaagctc atcgctttgc ggcagtagcg tttagagatg ctcaggtcaa
     1141 gcagcttaat aaccagccct ggcaaaccat aaaaaataca ctcacgcata acgggcatca
     1201 ctataccaac acgcagctcc ctgcagcaga gatgaaaatc ggcgcaaaag atatctttcc
     1261 cagtgcttat gagggaaagg gcgtatgcag ttgggatacc aagaatattc atcacgccaa
     1321 taatttgtgg atgtccacgg tgagtgtgca tgaggacggt aaagataaaa cgcttttttg
     1381 cgggatacgt catggcgtgc tttcccccta tcatgaaaaa gatccgcttc tgcgtcacgt
     1441 cggcgctgaa aacaaagcca aagaagtatt aactgcggca ctttttagta aacctgagtt
     1501 gcttaacaaa gccttagcgg gcgaggcggt aagcctgaaa ctggtatccg tcgggttact
     1561 caccgcgtcg aatattttcg gcaaagaggg aacgatggtc gaggaccaaa tgcgcgcatg
     1621 gcaatcgttg acccagccgg gaaaaatgat tcatttaaaa atccgcaata aagatggcga
     1681 tctacagacg gtaaaaataa aaccggacgt cgccgcattt aatgtgggtg ttaatgagct
     1741 ggcgctcaag ctcggctttg gccttaaggc atcggatagc tataatgccg aggcgctaca
     1801 tcagttatta ggcaatgatt tacgccctga agccagacca ggtggctggg ttggcgaatg
     1861 gctggcgcaa tacccggata attatgaggt cgtcaataca ttagcgcgcc agattaagga
     1921 tatatggaaa aataaccaac atcataaaga tggcggcgaa ccctataaac tcgcacaacg
     1981 ccttgccatg ttagcccatg aaattgacgc ggtacccgcc tggaattgta aaagcggcaa
     2041 agatcgtaca gggatgatgg attcagaaat caagcgagag atcatttcct tacatcagac
     2101 ccatatgtta agtgcgcctg gtagtcttcc ggatagcggt ggacagaaaa ttttccaaaa
     2161 agtattactg aatagcggta acctggagat tcagaaacaa aatacgggcg gggcgggaaa
     2221 caaagtaatg aaaaatttat cgccagaggt gctcaatctt tcctatcaaa aacgagttgg
     2281 ggatgaaaat atttggcagt cagtaaaagg catttcttca ttaatcacat ctcccgggat
     2341 ccaccggccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2401 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2461 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2521 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2581 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2641 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2701 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2761 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2821 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2881 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2941 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     3001 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3061 ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
     3121 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3181 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3241 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3301 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     3361 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3421 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3481 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3541 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3601 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3661 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3721 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3781 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3841 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3901 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4021 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4081 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4141 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4201 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4261 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4321 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4381 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4441 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4501 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4561 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4621 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4681 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4741 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4801 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4861 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4921 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4981 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5041 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5101 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5161 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5221 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5281 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5341 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5401 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5461 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5521 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5581 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5641 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5701 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5761 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5821 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5881 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5941 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6001 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6061 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6361 ttctttcctg cgttatcccc tgattctgtg gataaccgta t
//
