LOCUS       sequence-360249-        6198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminally tagged Salmonella Typhimurium SopB truncation for 
            mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6198)
  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 6198)
  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: EGFP-SopB(68-554)
FEATURES             Location/Qualifiers
     source          1..6198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(625..627,628..630,631..1341)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(625..627,628..630,631..1341)
                     /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(670..691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(931..950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1278..1299
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2998..3119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3035..3054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3089..3108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3126..3581)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3263..3284)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3475..3494
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3608..3712
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3714..4071
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3714..3905
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3737..3757)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3922..4057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4106..4900
                     /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             4106..4900
                     /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(4160..4179)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4770..4789
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5088..5107)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5132..5179
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5508..6096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5997..6016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      661 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      721 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      781 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      841 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      901 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      961 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1021 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1081 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1141 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1201 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1261 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1321 ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca
     1381 gtcgacggtg gtagtgcttc cggagcgtct aaccactcgc tgcataacct ctataactta
     1441 cagcgcgatc ttcttaccgt cgcggcaacc gttctgggta aacaagaccc ggttctaacg
     1501 tcaatggcaa accaaatgga gttagccaaa gttaaagcgg accggccagc aacaaaacaa
     1561 gaagaagccg cggcaaaagc attgaagaaa aatcttatcg aacttattgc agcacgcact
     1621 cagcagcagg atggcttacc tgcaaaagaa gctcatcgct ttgcggcagt agcgtttaga
     1681 gatgctcagg tcaagcagct taataaccag ccctggcaaa ccataaaaaa tacactcacg
     1741 cataacgggc atcactatac caacacgcag ctccctgcag cagagatgaa aatcggcgca
     1801 aaagatatct ttcccagtgc ttatgaggga aagggcgtat gcagttggga taccaagaat
     1861 attcatcacg ccaataattt gtggatgtcc acggtgagtg tgcatgagga cggtaaagat
     1921 aaaacgcttt tttgcgggat acgtcatggc gtgctttccc cctatcatga aaaagatccg
     1981 cttctgcgtc acgtcggcgc tgaaaacaaa gccaaagaag tattaactgc ggcacttttt
     2041 agtaaacctg agttgcttaa caaagcctta gcgggcgagg cggtaagcct gaaactggta
     2101 tccgtcgggt tactcaccgc gtcgaatatt ttcggcaaag agggaacgat ggtcgaggac
     2161 caaatgcgcg catggcaatc gttgacccag ccgggaaaaa tgattcattt aaaaatccgc
     2221 aataaagatg gcgatctaca gacggtaaaa ataaaaccgg acgtcgccgc atttaatgtg
     2281 ggtgttaatg agctggcgct caagctcggc tttggcctta aggcatcgga tagctataat
     2341 gccgaggcgc tacatcagtt attaggcaat gatttacgcc ctgaagccag accaggtggc
     2401 tgggttggcg aatggctggc gcaatacccg gataattatg aggtcgtcaa tacattagcg
     2461 cgccagatta aggatatatg gaaaaataac caacatcata aagatggcgg cgaaccctat
     2521 aaactcgcac aacgccttgc catgttagcc catgaaattg acgcggtacc cgcctggaat
     2581 tgtaaaagcg gcaaagatcg tacagggatg atggattcag aaatcaagcg agagatcatt
     2641 tccttacatc agacccatat gttaagtgcg cctggtagtc ttccggatag cggtggacag
     2701 aaaattttcc aaaaagtatt actgaatagc ggtaacctgg agattcagaa acaaaatacg
     2761 ggcggggcgg gaaacaaagt aatgaaaaat ttatcgccag aggtgctcaa tctttcctat
     2821 caaaaacgag ttggggatga aaatatttgg cagtcagtaa aaggctaagg atccaccgga
     2881 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2941 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3001 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3061 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3121 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3181 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3241 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3301 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3361 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3421 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3481 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3541 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3721 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3901 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3961 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4021 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4081 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4141 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4201 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4261 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4321 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4381 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4441 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4501 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4561 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4621 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4681 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4741 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4801 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4861 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4921 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4981 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5041 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5101 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5161 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5221 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5281 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5341 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5401 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5461 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5521 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5581 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5641 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5701 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5761 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5821 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5881 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5941 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6001 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6061 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6121 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6181 tctgtggata accgtatt
//
