LOCUS       sequence-179923-        8247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST tagged Munc18c with a F119E mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8247)
  AUTHORS   Latham CF, Lopez JA, Hu SH, Gee CL, Westbury E, Blair DH, Armishaw 
            CJ, Alewood PF, Bryant NJ, James DE, Martin JL
  TITLE     Molecular dissection of the Munc18c/syntaxin4 interaction: 
            implications for regulation of membrane trafficking.
  JOURNAL   Traffic. 2006 Oct;7(10):1408-19. Epub 2006 Aug 10.
  PUBMED    16899085
REFERENCE   2  (bases 1 to 8247)
  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: "Traffic."; 
            date: "2006-10"; volume: "7(10)"; pages: "1408-19. Epub 2006 Aug 10"
COMMENT     Sequence Label: Munc18cF119E-pDEST27
FEATURES             Location/Qualifiers
     source          1..8247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     761..778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(852..956)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1066..1369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1370..1573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1524..1544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1570..1594
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1683..2336
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2294..2316
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    2364..2388
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4171..4195)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4227..4246)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4228..4246)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4257..4274)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4257..4273)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4266..4288)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    4678..4812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4715..4734)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4769..4788
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4938..5393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5025..5044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5235..5256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5510..5867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5510..5701
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5533..5553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5718..5853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5918..6712
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5972..5991)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6582..6601
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6776..6824
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6865..6884)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6984..7006
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        7130..7234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7235..7303,7304..8095)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7453..7472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      781 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      841 cagagcttgc aattcgcgct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      961 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1021 aaaaataggc gtattacgag gccctttcac gcattagatg catgtcgtta cataacttac
     1081 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1141 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1201 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     1261 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1321 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1381 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1441 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1501 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1561 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     1621 tgacctccat agaagacacc gggaccgatc cagcctccgg actctagcct aggccgcgga
     1681 ccatggcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc
     1741 ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata
     1801 aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg
     1861 atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca
     1921 acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt
     1981 tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag
     2041 ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata
     2101 aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg
     2161 atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta
     2221 aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag
     2281 catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg
     2341 atctggttcc gcgttctaga tcaacaagtt tgtacaaaaa agcaggctta gatatggcgc
     2401 cgccggtatc ggagcgggga ctgaagagcg tcgtgtggcg gaagataaaa acagccgtgt
     2461 tcgatgactg ccggaaagaa ggcgaatgga agataatgct gttagatgag tttaccacca
     2521 aacttttgtc gtcatgctgc aaaatgacag accttctaga ggagggcata actgttatag
     2581 agaatattta taagaatcgt gaacctgtca gacaaatgaa agctctttat ttcatctctc
     2641 caacaccaaa atctgtagat tgtttcttga gagattttgg aagtaaatct gagaaaaaat
     2701 acaaagcagc gtatatatac ttcactgact tttgtcctga cagtctcgag aacaagatta
     2761 aagcatcttg ctccaagtca ataagaagat gtaaggaaat aaacatttcc ttcattccac
     2821 aggaatctca ggtttatact cttgatgtac cggacgcatt ctattactgt tacagtccag
     2881 accctagtaa cgccagcagg aaagaagtgg tcatggaggc aatggctgag cagattgtga
     2941 cagtgtgtgc cactctggat gaaaaccctg gagtgaggta caagagtaaa cctctagata
     3001 atgccagtaa gcttgcacag ctggttgaga aaaagcttga agactactac aaaattgatg
     3061 aaaaaggcct aataaagggt aaaactcagt cccagctctt aataattgac cgtggctttg
     3121 accctgtgtc cactgtcctg catgaactga cctttcaggc aatggcatat gatctactac
     3181 caattgagaa tgatacatac aagtacaaaa cagatggaaa agagaaggag gcagttcttg
     3241 aagaagacga cgacctgtgg gtgcgggttc gacaccggca catcgcggtt gtgttggagg
     3301 aaattccaaa gcttatgaag gaaatttcat caacaaagaa agctacagag gggaagacat
     3361 cacttagcgc tcttacccag ctgatgaaaa agatgccgca cttccgaaag cagatctcga
     3421 agcaagtagt ccatcttaac ttagctgaag actgcatgaa taagtttaag ctgaatattg
     3481 agaagctctg caaaactgag caggacctgg cacttggaac agatgctgaa ggccagcggg
     3541 tgaaggactc catgctggtg ctcctcccag tgctgctcaa caaaaaccat gacaactgcg
     3601 acaaaatacg ggcagtcctg ctctacatct tcgggattaa tggaaccact gaagaaaatc
     3661 tggacagact gatccacaat gtaaagatag aagatgatag tgatatgatt cgtaactgga
     3721 gccaccttgg tgttcccatt gttcccccat cccagcaagc caaaccactg agaaaggatc
     3781 ggtctgcaga agagactttt cagctttctc gatggacacc ttttatcaaa gatatcatgg
     3841 aggatgccat tgataataga ttagattcca aagagtggcc gtattgttcc cggtgcccag
     3901 cagtgtggaa tggctctgga gctgtgagtg ctcgccagaa acccagaact aactacttag
     3961 agctggaccg gaaaaatggg tcaaggctga ttatttttgt aattggagga attacgtact
     4021 ctgagatgcg gtgtgcttat gaagtttccc aggcacataa atcctgtgag gttattattg
     4081 gttccacaca tattttaaca cccagaaagc tcctggatga tataaaaatg ctgaataaat
     4141 caaaggataa agtttccttt aaggatgagg acccagcttt cttgtacaaa gtggttgatc
     4201 gcgtgcatgc gacgtcatag ctctctccct atagtgagtc gtattataag ctaggcactg
     4261 gccgtcgttt tacaacgtcg tgactgggaa aactgctagc ttgggatctt tgtgaaggaa
     4321 ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag
     4381 gtaaatataa aatttttaag tgtataatgt gttaaactag ctgcatatgc ttgctgcttg
     4441 agagttttgc ttactgagta tgatttatga aaatattata cacaggagct agtgattcta
     4501 attgtttgtg tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac
     4561 agtctgttca tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4621 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4681 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4801 catgtctgga tcgatcctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4861 tattggctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4921 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5341 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     5401 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag
     5461 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa
     5521 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgattctt
     5881 ctgacacaac agtctcgaac ttaaggctag agccaccatg attgaacaag atggattgca
     5941 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     6001 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     6061 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     6121 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     6181 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6241 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     6301 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     6361 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6421 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     6481 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6541 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6601 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6661 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6721 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     6781 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     6841 gatccgcgta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6901 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6961 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     7021 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     7081 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     7141 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     7201 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7261 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7321 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7381 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7441 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7501 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7561 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7621 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7681 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7741 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7801 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7861 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7921 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7981 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     8041 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     8101 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8161 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8221 gttccactga gcgtcagacc ccgtaga
//
