LOCUS       sequence-360254-        5769 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 5769)
  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 5769)
  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(211-554)
FEATURES             Location/Qualifiers
     source          1..5769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..353
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        354..557
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     508..528
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      596..605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(602..604,605..607,608..1318)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(602..604,605..607,608..1318)
                     /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(647..668)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(908..927)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1255..1276
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2546..2667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2583..2602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2637..2656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2674..3129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3023..3042
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3156..3260
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3262..3619
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3262..3453
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3285..3305)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3470..3605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3654..4448
                     /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             3654..4448
                     /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(3708..3727)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4318..4337
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4636..4655)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4680..4727
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5056..5644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5545..5564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
       61 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      121 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      181 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      241 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      301 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      361 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      421 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      481 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      541 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac
      601 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      661 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      721 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      781 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      841 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      901 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      961 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1021 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1081 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1141 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1201 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1261 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     1321 cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtggta gtgcttccgg
     1381 aggcgtatgc agttgggata ccaagaatat tcatcacgcc aataatttgt ggatgtccac
     1441 ggtgagtgtg catgaggacg gtaaagataa aacgcttttt tgcgggatac gtcatggcgt
     1501 gctttccccc tatcatgaaa aagatccgct tctgcgtcac gtcggcgctg aaaacaaagc
     1561 caaagaagta ttaactgcgg cactttttag taaacctgag ttgcttaaca aagccttagc
     1621 gggcgaggcg gtaagcctga aactggtatc cgtcgggtta ctcaccgcgt cgaatatttt
     1681 cggcaaagag ggaacgatgg tcgaggacca aatgcgcgca tggcaatcgt tgacccagcc
     1741 gggaaaaatg attcatttaa aaatccgcaa taaagatggc gatctacaga cggtaaaaat
     1801 aaaaccggac gtcgccgcat ttaatgtggg tgttaatgag ctggcgctca agctcggctt
     1861 tggccttaag gcatcggata gctataatgc cgaggcgcta catcagttat taggcaatga
     1921 tttacgccct gaagccagac caggtggctg ggttggcgaa tggctggcgc aatacccgga
     1981 taattatgag gtcgtcaata cattagcgcg ccagattaag gatatatgga aaaataacca
     2041 acatcataaa gatggcggcg aaccctataa actcgcacaa cgccttgcca tgttagccca
     2101 tgaaattgac gcggtacccg cctggaattg taaaagcggc aaagatcgta cagggatgat
     2161 ggattcagaa atcaagcgag agatcatttc cttacatcag acccatatgt taagtgcgcc
     2221 tggtagtctt ccggatagcg gtggacagaa aattttccaa aaagtattac tgaatagcgg
     2281 taacctggag attcagaaac aaaatacggg cggggcggga aacaaagtaa tgaaaaattt
     2341 atcgccagag gtgctcaatc tttcctatca aaaacgagtt ggggatgaaa atatttggca
     2401 gtcagtaaaa ggctaaggat ccaccggatc tagataactg atcataatca gccataccac
     2461 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2521 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2641 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     2701 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2761 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2821 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2881 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2941 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3001 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3061 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3121 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3181 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3241 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3301 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3421 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3481 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3541 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3601 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3661 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3721 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3781 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3841 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3901 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3961 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4021 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4081 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4141 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4201 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4261 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4321 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4381 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4441 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4501 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4561 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4621 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4681 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4741 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4801 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4861 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4921 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4981 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5041 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5101 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5161 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5221 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5281 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5341 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5401 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5461 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5521 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5581 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5641 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5701 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     5761 gttattaat
//
