LOCUS       sequence-360242-        6303 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 6303)
  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 6303)
  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(33-554)
FEATURES             Location/Qualifiers
     source          1..6303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      612..621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(618..620,621..623,624..1334)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(618..620,621..623,624..1334)
                     /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(663..684)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(924..943)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1271..1292
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3096..3217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3133..3152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3187..3206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3224..3679)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3361..3382)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3573..3592
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3706..3810
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3812..4169
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3812..4003
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3835..3855)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4020..4155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4204..4998
                     /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             4204..4998
                     /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(4258..4277)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4868..4887
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5186..5205)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5230..5277
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5606..6194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6095..6114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      601 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      661 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      721 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      781 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
      841 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      901 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      961 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1021 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1081 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1141 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1201 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     1261 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1321 acgagctgta caagtccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
     1381 gtggtagtgc ttccggatca ggccagggca aagcgccggc taaagcgccc gacgctcgcc
     1441 cggaaattat tgtcctgcga gaacccggcg cgacatgggg gaattatcta cagcatcaga
     1501 aggcgtctaa ccactcgctg cataacctct ataacttaca gcgcgatctt cttaccgtcg
     1561 cggcaaccgt tctgggtaaa caagacccgg ttctaacgtc aatggcaaac caaatggagt
     1621 tagccaaagt taaagcggac cggccagcaa caaaacaaga agaagccgcg gcaaaagcat
     1681 tgaagaaaaa tcttatcgaa cttattgcag cacgcactca gcagcaggat ggcttacctg
     1741 caaaagaagc tcatcgcttt gcggcagtag cgtttagaga tgctcaggtc aagcagctta
     1801 ataaccagcc ctggcaaacc ataaaaaata cactcacgca taacgggcat cactatacca
     1861 acacgcagct ccctgcagca gagatgaaaa tcggcgcaaa agatatcttt cccagtgctt
     1921 atgagggaaa gggcgtatgc agttgggata ccaagaatat tcatcacgcc aataatttgt
     1981 ggatgtccac ggtgagtgtg catgaggacg gtaaagataa aacgcttttt tgcgggatac
     2041 gtcatggcgt gctttccccc tatcatgaaa aagatccgct tctgcgtcac gtcggcgctg
     2101 aaaacaaagc caaagaagta ttaactgcgg cactttttag taaacctgag ttgcttaaca
     2161 aagccttagc gggcgaggcg gtaagcctga aactggtatc cgtcgggtta ctcaccgcgt
     2221 cgaatatttt cggcaaagag ggaacgatgg tcgaggacca aatgcgcgca tggcaatcgt
     2281 tgacccagcc gggaaaaatg attcatttaa aaatccgcaa taaagatggc gatctacaga
     2341 cggtaaaaat aaaaccggac gtcgccgcat ttaatgtggg tgttaatgag ctggcgctca
     2401 agctcggctt tggccttaag gcatcggata gctataatgc cgaggcgcta catcagttat
     2461 taggcaatga tttacgccct gaagccagac caggtggctg ggttggcgaa tggctggcgc
     2521 aatacccgga taattatgag gtcgtcaata cattagcgcg ccagattaag gatatatgga
     2581 aaaataacca acatcataaa gatggcggcg aaccctataa actcgcacaa cgccttgcca
     2641 tgttagccca tgaaattgac gcggtacccg cctggaattg taaaagcggc aaagatcgta
     2701 cagggatgat ggattcagaa atcaagcgag agatcatttc cttacatcag acccatatgt
     2761 taagtgcgcc tggtagtctt ccggatagcg gtggacagaa aattttccaa aaagtattac
     2821 tgaatagcgg taacctggag attcagaaac aaaatacggg cggggcggga aacaaagtaa
     2881 tgaaaaattt atcgccagag gtgctcaatc tttcctatca aaaacgagtt ggggatgaaa
     2941 atatttggca gtcagtaaaa ggctaaggat ccaccggatc tagataactg atcataatca
     3001 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3061 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3121 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3181 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     3241 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3301 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3361 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3421 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3481 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3541 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3601 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3661 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3781 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3841 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3901 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3961 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4021 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4081 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4141 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4201 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4261 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4321 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4381 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4441 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4501 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4561 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4621 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4681 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4741 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4801 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4861 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4921 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4981 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5041 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5101 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5161 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5221 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5281 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5341 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5401 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5461 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5521 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6301 cca
//
