LOCUS       sequence-179924-        8247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST tagged Munc18c.
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: Munc18cwt-pDEST27
FEATURES             Location/Qualifiers
     source          1..8247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(835..939)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1049..1352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1353..1556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1507..1527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1553..1577
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1666..2319
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2277..2299
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    2347..2371
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4154..4178)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4210..4229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4211..4229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4240..4257)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4240..4256)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4249..4271)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    4661..4795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4698..4717)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4752..4771
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4921..5376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5008..5027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5218..5239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5493..5850
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5493..5684
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5516..5536)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5701..5836
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5901..6695
                     /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(5955..5974)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6565..6584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6759..6807
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6848..6867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6967..6989
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        7113..7217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7218..7286,7287..8078)
                     /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(7436..7455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagagct tgcaattcgc
      841 gctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      961 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtattac
     1021 gaggcccttt cacgcattag atgcatgtcg ttacataact tacggtaaat ggcccgcctg
     1081 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     1141 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     1201 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     1261 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     1321 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     1381 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     1441 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     1501 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     1561 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
     1621 accgggaccg atccagcctc cggactctag cctaggccgc ggaccatggc ccctatacta
     1681 ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa
     1741 gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag
     1801 tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta
     1861 acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt
     1921 ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt
     1981 gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag
     2041 ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt
     2101 gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg
     2161 gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct
     2221 atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc
     2281 tggcaagcca cgtttggtgg tggcgaccat cctccaaaat cggatctggt tccgcgttct
     2341 agatcaacaa gtttgtacaa aaaagcaggc ttagatatgg cgccgccggt atcggagcgg
     2401 ggactgaaga gcgtcgtgtg gcggaagata aaaacagccg tgttcgatga ctgccggaaa
     2461 gaaggcgaat ggaagataat gctgttagat gagtttacca ccaaactttt gtcgtcatgc
     2521 tgcaaaatga cagaccttct agaggagggc ataactgtta tagagaatat ttataagaat
     2581 cgtgaacctg tcagacaaat gaaagctctt tatttcatct ctccaacacc aaaatctgta
     2641 gattgtttct tgagagattt tggaagtaaa tctgagaaaa aatacaaagc agcgtatata
     2701 tacttcactg acttttgtcc tgacagtctc tttaacaaga ttaaagcatc ttgctccaag
     2761 tcaataagaa gatgtaagga aataaacatt tccttcattc cacaggaatc tcaggtttat
     2821 actcttgatg taccggacgc attctattac tgttacagtc cagaccctag taacgccagc
     2881 aggaaagaag tggtcatgga ggcaatggct gagcagattg tgacagtgtg tgccactctg
     2941 gatgaaaacc ctggagtgag gtacaagagt aaacctctag ataatgccag taagcttgca
     3001 cagctggttg agaaaaagct tgaagactac tacaaaattg atgaaaaagg cctaataaag
     3061 ggtaaaactc agtcccagct cttaataatt gaccgtggct ttgaccctgt gtccactgtc
     3121 ctgcatgaac tgacctttca ggcaatggca tatgatctac taccaattga gaatgataca
     3181 tacaagtaca aaacagatgg aaaagagaag gaggcagttc ttgaagaaga cgacgacctg
     3241 tgggtgcggg ttcgacaccg gcacatcgcg gttgtgttgg aggaaattcc aaagcttatg
     3301 aaggaaattt catcaacaaa gaaagctaca gaggggaaga catcacttag cgctcttacc
     3361 cagctgatga aaaagatgcc gcacttccga aagcagatct cgaagcaagt agtccatctt
     3421 aacttagctg aagactgcat gaataagttt aagctgaata ttgagaagct ctgcaaaact
     3481 gagcaggacc tggcacttgg aacagatgct gaaggccagc gggtgaagga ctccatgctg
     3541 gtgctcctcc cagtgctgct caacaaaaac catgacaact gcgacaaaat acgggcagtc
     3601 ctgctctaca tcttcgggat taatggaacc actgaagaaa atctggacag actgatccac
     3661 aatgtaaaga tagaagatga tagtgatatg attcgtaact ggagccacct tggtgttccc
     3721 attgttcccc catcccagca agccaaacca ctgagaaagg atcggtctgc agaagagact
     3781 tttcagcttt ctcgatggac accttttatc aaagatatca tggaggatgc cattgataat
     3841 agattagatt ccaaagagtg gccgtattgt tcccggtgcc cagcagtgtg gaatggctct
     3901 ggagctgtga gtgctcgcca gaaacccaga actaactact tagagctgga ccggaaaaat
     3961 gggtcaaggc tgattatttt tgtaattgga ggaattacgt actctgagat gcggtgtgct
     4021 tatgaagttt cccaggcaca taaatcctgt gaggttatta ttggttccac acatatttta
     4081 acacccagaa agctcctgga tgatataaaa atgctgaata aatcaaagga taaagtttcc
     4141 tttaaggatg aggacccagc tttcttgtac aaagtggttg atcgcgtgca tgcgacgtca
     4201 tagctctctc cctatagtga gtcgtattat aagctaggca ctggccgtcg ttttacaacg
     4261 tcgtgactgg gaaaactgct agcttgggat ctttgtgaag gaaccttact tctgtggtgt
     4321 gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt
     4381 aagtgtataa tgtgttaaac tagctgcata tgcttgctgc ttgagagttt tgcttactga
     4441 gtatgattta tgaaaatatt atacacagga gctagtgatt ctaattgttt gtgtatttta
     4501 gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt tcatgatcat
     4561 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     4621 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     4681 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4741 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgatcc
     4801 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc tggcgtaata
     4861 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     4921 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4981 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5041 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5101 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     5161 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5221 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     5281 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5341 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     5401 gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata
     5461 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     5521 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5581 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5641 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5701 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5761 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5821 ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg
     5881 aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5941 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     6001 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     6061 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     6121 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6181 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6241 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6301 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6361 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6421 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6481 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6541 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6601 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6661 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6721 cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat
     6781 tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca
     6841 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     6901 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6961 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     7021 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     7081 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7141 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7201 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7261 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7321 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7381 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7441 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7501 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7561 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     7621 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     7681 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7741 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7801 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7861 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7921 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7981 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8041 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8101 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8161 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8221 accccgtaga aaagatcaaa ggatctt
//
