LOCUS       sequence-360229-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-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 6401)
  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 6401)
  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: SopB(C460S)-EGFP
FEATURES             Location/Qualifiers
     source          1..6401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        33..336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        337..540
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     491..511
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2313..2322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2319..2321,2322..2324,2325..3038)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2319..2321,2322..2324,2325..3035)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2364..2385)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2625..2644)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2972..2993
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3161..3282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3198..3217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3252..3271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3289..3744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3426..3447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3638..3657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3771..3875
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3877..4234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3877..4068
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3900..3920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4269..5063
                     /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             4269..5063
                     /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(4323..4342)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4933..4952
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5251..5270)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5295..5342
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5671..6259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6160..6179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
       61 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      121 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      181 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      241 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      301 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      361 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      421 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      481 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      541 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
      601 gaattctaat gcaaatacag agcttctatc actcagcttc actaaaaacc caggaggctt
      661 ttaaaagcct acaaaaaacc ttatacaacg gaatgcagat tctctcaggc cagggcaaag
      721 cgccggctaa agcgcccgac gctcgcccgg aaattattgt cctgcgagaa cccggcgcga
      781 catgggggaa ttatctacag catcagaagg cgtctaacca ctcgctgcat aacctctata
      841 acttacagcg cgatcttctt accgtcgcgg caaccgttct gggtaaacaa gacccggttc
      901 taacgtcaat ggcaaaccaa atggagttag ccaaagttaa agcggaccgg ccagcaacaa
      961 aacaagaaga agccgcggca aaagcattga agaaaaatct tatcgaactt attgcagcac
     1021 gcactcagca gcaggatggc ttacctgcaa aagaagctca tcgctttgcg gcagtagcgt
     1081 ttagagatgc tcaggtcaag cagcttaata accagccctg gcaaaccata aaaaatacac
     1141 tcacgcataa cgggcatcac tataccaaca cgcagctccc tgcagcagag atgaaaatcg
     1201 gcgcaaaaga tatctttccc agtgcttatg agggaaaggg cgtatgcagt tgggatacca
     1261 agaatattca tcacgccaat aatttgtgga tgtccacggt gagtgtgcat gaggacggta
     1321 aagataaaac gcttttttgc gggatacgtc atggcgtgct ttccccctat catgaaaaag
     1381 atccgcttct gcgtcacgtc ggcgctgaaa acaaagccaa agaagtatta actgcggcac
     1441 tttttagtaa acctgagttg cttaacaaag ccttagcggg cgaggcggta agcctgaaac
     1501 tggtatccgt cgggttactc accgcgtcga atattttcgg caaagaggga acgatggtcg
     1561 aggaccaaat gcgcgcatgg caatcgttga cccagccggg aaaaatgatt catttaaaaa
     1621 tccgcaataa agatggcgat ctacagacgg taaaaataaa accggacgtc gccgcattta
     1681 atgtgggtgt taatgagctg gcgctcaagc tcggctttgg ccttaaggca tcggatagct
     1741 ataatgccga ggcgctacat cagttattag gcaatgattt acgccctgaa gccagaccag
     1801 gtggctgggt tggcgaatgg ctggcgcaat acccggataa ttatgaggtc gtcaatacat
     1861 tagcgcgcca gattaaggat atatggaaaa ataaccaaca tcataaagat ggcggcgaac
     1921 cctataaact cgcacaacgc cttgccatgt tagcccatga aattgacgcg gtacccgcct
     1981 ggaattctaa aagcggcaaa gatcgtacag ggatgatgga ttcagaaatc aagcgagaga
     2041 tcatttcctt acatcagacc catatgttaa gtgcgcctgg tagtcttccg gatagcggtg
     2101 gacagaaaat tttccaaaaa gtattactga atagcggtaa cctggagatt cagaaacaaa
     2161 atacgggcgg ggcgggaaac aaagtaatga aaaatttatc gccagaggtg ctcaatcttt
     2221 cctatcaaaa acgagttggg gatgaaaata tttggcagtc agtaaaaggc atttcttcat
     2281 taatcacatc tcccgggatc caccggccgg tcgccaccat ggtgagcaag ggcgaggagc
     2341 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2401 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2461 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2521 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2581 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2641 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2701 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2761 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2821 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2881 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     2941 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3001 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     3061 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3121 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3181 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3241 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3301 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3361 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3421 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3481 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3541 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3601 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3661 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3721 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3781 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3841 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3901 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4261 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4321 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4381 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4441 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4501 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4561 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4621 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4681 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4741 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4801 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4861 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4921 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4981 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5041 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5101 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5161 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5221 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5281 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5341 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5401 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5461 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6301 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6361 taccgccatg cattagttat taatagtaat caattacggg g
//
