LOCUS       sequence-360245-        6204 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 6204)
  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 6204)
  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(33-520)
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      609..618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(615..617,618..620,621..1331)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(615..617,618..620,621..1331)
                     /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(660..681)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(921..940)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1268..1289
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2994..3115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3031..3050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3085..3104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3122..3577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3259..3280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3471..3490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3604..3708
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3710..4067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3710..3901
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3733..3753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3918..4053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4102..4896
                     /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             4102..4896
                     /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(4156..4175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4766..4785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5084..5103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5128..5175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5504..6092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5993..6012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      601 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      661 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      721 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      781 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      841 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      901 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      961 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1021 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1081 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1141 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1201 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1261 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1321 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggtg
     1381 gtagtgcttc cggatcaggc cagggcaaag cgccggctaa agcgcccgac gctcgcccgg
     1441 aaattattgt cctgcgagaa cccggcgcga catgggggaa ttatctacag catcagaagg
     1501 cgtctaacca ctcgctgcat aacctctata acttacagcg cgatcttctt accgtcgcgg
     1561 caaccgttct gggtaaacaa gacccggttc taacgtcaat ggcaaaccaa atggagttag
     1621 ccaaagttaa agcggaccgg ccagcaacaa aacaagaaga agccgcggca aaagcattga
     1681 agaaaaatct tatcgaactt attgcagcac gcactcagca gcaggatggc ttacctgcaa
     1741 aagaagctca tcgctttgcg gcagtagcgt ttagagatgc tcaggtcaag cagcttaata
     1801 accagccctg gcaaaccata aaaaatacac tcacgcataa cgggcatcac tataccaaca
     1861 cgcagctccc tgcagcagag atgaaaatcg gcgcaaaaga tatctttccc agtgcttatg
     1921 agggaaaggg cgtatgcagt tgggatacca agaatattca tcacgccaat aatttgtgga
     1981 tgtccacggt gagtgtgcat gaggacggta aagataaaac gcttttttgc gggatacgtc
     2041 atggcgtgct ttccccctat catgaaaaag atccgcttct gcgtcacgtc ggcgctgaaa
     2101 acaaagccaa agaagtatta actgcggcac tttttagtaa acctgagttg cttaacaaag
     2161 ccttagcggg cgaggcggta agcctgaaac tggtatccgt cgggttactc accgcgtcga
     2221 atattttcgg caaagaggga acgatggtcg aggaccaaat gcgcgcatgg caatcgttga
     2281 cccagccggg aaaaatgatt catttaaaaa tccgcaataa agatggcgat ctacagacgg
     2341 taaaaataaa accggacgtc gccgcattta atgtgggtgt taatgagctg gcgctcaagc
     2401 tcggctttgg ccttaaggca tcggatagct ataatgccga ggcgctacat cagttattag
     2461 gcaatgattt acgccctgaa gccagaccag gtggctgggt tggcgaatgg ctggcgcaat
     2521 acccggataa ttatgaggtc gtcaatacat tagcgcgcca gattaaggat atatggaaaa
     2581 ataaccaaca tcataaagat ggcggcgaac cctataaact cgcacaacgc cttgccatgt
     2641 tagcccatga aattgacgcg gtacccgcct ggaattgtaa aagcggcaaa gatcgtacag
     2701 ggatgatgga ttcagaaatc aagcgagaga tcatttcctt acatcagacc catatgttaa
     2761 gtgcgcctgg tagtcttccg gatagcggtg gacagaaaat tttccaaaaa gtattactga
     2821 atagcggtaa cctggagatt cagaaacaaa atacgggcgg gtaaggatcc accggatcta
     2881 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2941 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3001 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3061 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     3121 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3181 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3241 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3301 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3361 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3421 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3481 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3541 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3601 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3661 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3721 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4081 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4141 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4201 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4261 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4321 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4381 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4441 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4501 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4561 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4621 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4681 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4741 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4801 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4861 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4921 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4981 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5041 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5101 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5161 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5221 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5281 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5341 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5401 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5461 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5521 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5581 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5641 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5701 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5761 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5821 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5881 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5941 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6001 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6061 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6121 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6181 tggataaccg tattaccgcc atgc
//
