LOCUS       sequence-362579-        5544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Legionella pneumophila SidK (1-278) - GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5544)
  AUTHORS   Maxson ME, Abbas YM, Wu JZ, Plumb JD, Grinstein S, Rubinstein JL
  TITLE     Detection and quantification of the vacuolar H+ATPase using the 
            Legionella effector protein SidK.
  JOURNAL   J Cell Biol. 2022 Mar 7;221(3). pii: 212963. doi: 
            10.1083/jcb.202107174. Epub 2022 Jan 13.
  PUBMED    35024770
REFERENCE   2  (bases 1 to 5544)
  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: "J Cell 
            Biol."; date: "2022-03-7"; volume: "221(3). pii: 212963. doi"; 
            pages: " 10.1083/jcb.202107174. Epub 2022 Jan 13"
COMMENT     Sequence Label: pYMA3_SidK_GFP
FEATURES             Location/Qualifiers
     source          1..5544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..151
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1096..1105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1102..1104,1105..1107,1108..1821)
                     /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(1102..1104,1105..1107,1108..1818)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1147..1168)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1408..1427)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1755..1776
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1944..2065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1981..2000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2035..2054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2072..2527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2209..2230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2421..2440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2554..2658
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2660..3017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2660..2851
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2683..2703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2868..3003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3052..3846
                     /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             3052..3846
                     /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(3106..3125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3716..3735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4034..4053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4078..4125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4454..5042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4943..4962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5217..5520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5521..5544,1..180)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
       61 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      121 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      181 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
      241 gaattcatgt cttttatcaa ggtaggtata aaaatgggtg gcttaactag cgagcaatat
      301 catagtcaag tagtcggtaa aatcggatat atcgccagat gcatgcaaac tatagatcct
      361 gaaaataatt tgaaaaaaat aagggaagat taccaagatg tattgatatg ggcagagaaa
      421 aattatcgtt ttgaggaaat attggaagcc agcaaatctg gaaaatgccc taatgatctg
      481 gatgcattaa gtcgacgatc gctaatactc caagaattgc tgcggcttgt ttcatcaatt
      541 agtcccttta aaatgaaatt agacctgatt gagtctcaat atgagaaaat gaagcagcat
      601 gtgaatttat ggaaaagtga ttaccatgta aaactaaatc aactgaatca attaacagat
      661 tacctcaaaa atgcagcacc aactcctaaa aataattttc taagagcaat gacctctgta
      721 ttgcagatgc agatagccca atatgggatt acagaagata atgaaggcat caaccagttg
      781 tttaaactgg gattgcattt gctcgcaatg gctaacgaaa aaatagatga acaatatcac
      841 ctgtttaaag ggtatgtaaa agatcagcct gaagaatctc cttttgaagg tattttgcct
      901 gcggaagacc agaaaatttt agttaaaact atgatagatt atgcaatgcc gaagctttca
      961 agcaaggttt tacaagataa attatcggca ctatccagtt ctgatgtatt aacaaaaaca
     1021 ttacttgata gcatcgacag aattgttaaa gaaaacgaaa aattaaatgc tttaagcaaa
     1081 ggggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1141 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1201 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1261 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1321 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1381 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1441 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1501 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1561 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1621 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1681 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1741 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1801 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     1861 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1921 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1981 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2041 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     2101 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2161 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2221 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2281 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2341 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2401 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2461 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2521 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2581 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2641 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2701 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2761 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2881 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2941 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3001 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3061 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3121 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3181 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3241 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3301 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3361 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3421 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3481 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3541 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     3601 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3661 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3721 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3781 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3841 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3901 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3961 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4021 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4081 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4141 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4201 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4261 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4321 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4381 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4441 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4501 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4561 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4621 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4681 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4741 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4801 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4861 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4921 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4981 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5041 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5101 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     5161 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5221 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     5281 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     5341 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     5401 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     5461 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     5521 gtgatgcggt tttggcagta catc
//
