LOCUS       sequence-360234-        6417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminally tagged Salmonella Typhimurium SopB (catalytically 
            inactive) for mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6417)
  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 6417)
  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(C460S)
FEATURES             Location/Qualifiers
     source          1..6417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        375..578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     529..549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      617..626
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(623..625,626..628,629..1339)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(623..625,626..628,629..1339)
                     /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(668..689)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(929..948)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1276..1297
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3215..3336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3252..3271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3306..3325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3343..3798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3480..3501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3692..3711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3825..3929
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3931..4288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3931..4122
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3954..3974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4139..4274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4323..5117
                     /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             4323..5117
                     /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(4377..4396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4987..5006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5305..5324)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5349..5396
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5725..6313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6214..6233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
       61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      361 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      541 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      601 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      661 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      721 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      781 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      841 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      901 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      961 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1021 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1081 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1141 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1201 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1261 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1321 catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt
     1381 cgacggtggt agtgcttccg gacaaataca gagcttctat cactcagctt cactaaaaac
     1441 ccaggaggct tttaaaagcc tacaaaaaac cttatacaac ggaatgcaga ttctctcagg
     1501 ccagggcaaa gcgccggcta aagcgcccga cgctcgcccg gaaattattg tcctgcgaga
     1561 acccggcgcg acatggggga attatctaca gcatcagaag gcgtctaacc actcgctgca
     1621 taacctctat aacttacagc gcgatcttct taccgtcgcg gcaaccgttc tgggtaaaca
     1681 agacccggtt ctaacgtcaa tggcaaacca aatggagtta gccaaagtta aagcggaccg
     1741 gccagcaaca aaacaagaag aagccgcggc aaaagcattg aagaaaaatc ttatcgaact
     1801 tattgcagca cgcactcagc agcaggatgg cttacctgca aaagaagctc atcgctttgc
     1861 ggcagtagcg tttagagatg ctcaggtcaa gcagcttaat aaccagccct ggcaaaccat
     1921 aaaaaataca ctcacgcata acgggcatca ctataccaac acgcagctcc ctgcagcaga
     1981 gatgaaaatc ggcgcaaaag atatctttcc cagtgcttat gagggaaagg gcgtatgcag
     2041 ttgggatacc aagaatattc atcacgccaa taatttgtgg atgtccacgg tgagtgtgca
     2101 tgaggacggt aaagataaaa cgcttttttg cgggatacgt catggcgtgc tttcccccta
     2161 tcatgaaaaa gatccgcttc tgcgtcacgt cggcgctgaa aacaaagcca aagaagtatt
     2221 aactgcggca ctttttagta aacctgagtt gcttaacaaa gccttagcgg gcgaggcggt
     2281 aagcctgaaa ctggtatccg tcgggttact caccgcgtcg aatattttcg gcaaagaggg
     2341 aacgatggtc gaggaccaaa tgcgcgcatg gcaatcgttg acccagccgg gaaaaatgat
     2401 tcatttaaaa atccgcaata aagatggcga tctacagacg gtaaaaataa aaccggacgt
     2461 cgccgcattt aatgtgggtg ttaatgagct ggcgctcaag ctcggctttg gccttaaggc
     2521 atcggatagc tataatgccg aggcgctaca tcagttatta ggcaatgatt tacgccctga
     2581 agccagacca ggtggctggg ttggcgaatg gctggcgcaa tacccggata attatgaggt
     2641 cgtcaataca ttagcgcgcc agattaagga tatatggaaa aataaccaac atcataaaga
     2701 tggcggcgaa ccctataaac tcgcacaacg ccttgccatg ttagcccatg aaattgacgc
     2761 ggtacccgcc tggaattcta aaagcggcaa agatcgtaca gggatgatgg attcagaaat
     2821 caagcgagag atcatttcct tacatcagac ccatatgtta agtgcgcctg gtagtcttcc
     2881 ggatagcggt ggacagaaaa ttttccaaaa agtattactg aatagcggta acctggagat
     2941 tcagaaacaa aatacgggcg gggcgggaaa caaagtaatg aaaaatttat cgccagaggt
     3001 gctcaatctt tcctatcaaa aacgagttgg ggatgaaaat atttggcagt cagtaaaagg
     3061 catttcttca ttaatcacat cttaaggatc caccggatct agataactga tcataatcag
     3121 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3181 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3241 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3301 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3361 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3421 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3481 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3541 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3601 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3661 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3721 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3781 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3841 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3901 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3961 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4021 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4141 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4201 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4261 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4321 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4381 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4441 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4501 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4561 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4621 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4681 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4741 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4801 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4861 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4921 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4981 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5041 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5101 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5161 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5221 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5281 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5341 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5401 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5461 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5521 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5581 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5641 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattac
//
