LOCUS       sequence-272494-        6603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express rat Munc18b in Sf9 cells, the resulted plasmid encodes an 
            N-terminally His6-tagged Munc18c protein with a tobacco etch virus 
            (TEV) cleavage site between the His6 tag and Munc18b..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6603)
  AUTHORS   Yu H, Crisman L, Stowell MHB, Shen J
  TITLE     Functional Reconstitution of Intracellular Vesicle Fusion Using 
            Purified SNAREs and Sec1/Munc18 (SM) Proteins.
  JOURNAL   Methods Mol Biol. 2019;1860:237-249. doi: 
            10.1007/978-1-4939-8760-3_15.
  PUBMED    30317509
REFERENCE   2  (bases 1 to 6603)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "1860:237-249. doi"; pages: " 
            10.1007/978-1-4939-8760-3_15"
COMMENT     Sequence Label: pFastBac HT JS-Munc18b
FEATURES             Location/Qualifiers
     source          1..6603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..3533))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3885..3976
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3901..3918
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4043..4060
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4082..4102
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    6093..6227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6130..6149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6184..6203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6256..6421
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3961 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatctcg
     4021 gtccgaaacc atgtcgtact accatcacca tcaccatcac gattacgata tcccaacgac
     4081 cgaaaacctg tattttcagg gcgccatgga tccgatggcg cccttggggc tgaaggcggt
     4141 ggtgggggaa aaaattctaa gtggagtcat ccggagcgtt aagaaggatg gcgagtggaa
     4201 ggtactcatc atggatcatc cgagcatgcg aatcttgtca tcctgttgta aaatgtcaga
     4261 tatcttggcg gagggcatca ccattgtgga agacatcaac aaacggagag aacccattcc
     4321 cagcttggag gcaatttatt tgctgagtcc cacagagaag tcggttcagg ctctgattgc
     4381 ggacttccag gggacaccaa ccttcaccta caaagcagcc catatcttct tcacggacac
     4441 gtgccccgag cccttgttca gtgaactggg ccgctctcgc ctggcgaagg tggtgaagac
     4501 actgaaagag atccacctcg ccttcctccc ctatgaggcc caggtgttct ctctggatgc
     4561 cccgcacagc acctacaacc tttattgccc atttcgggca ggggagcggg ggcggcagct
     4621 cgatgtgctg gcccagcaga tagccacact gtgtgccact cttcaggaat acccatccat
     4681 tcgctaccgc aagggcccag aagacacagc ccagctggcc catgctgtcc tggccaagct
     4741 gaacgccttc aaggcagaca ctcccagtct gggcgagggc ccagagaaaa cacgctcaca
     4801 gttgctcata atggaccggg cagcagaccc cgtgtccccg ctattacatg aactcacatt
     4861 ccaagccatg gcctatgatc ttctgcacat cgagcaggac acatacaggt atgagaccac
     4921 aggactgagc gagtcccggg agaaggctgt gctcctggat gaagacgatg acctgtgggt
     4981 ggaactccgg cacatgcaca tagcggatgt gtccaagaag gtcacggagc tcctgaagac
     5041 attctgtgag agcaagcggc tgaccacaga caaggccaac atcaaagacc tgtcccacat
     5101 cctgaaaaag atgccacagt atcagaagga gttgaacaag tactccacgc acctgcactt
     5161 ggcagatgac tgcatgaagc acttcaaggg ctcggtggag aagctgtgcg gtgtggagca
     5221 ggacctggcc atgggctctg atgcagaggg tgagaagatc aaggacacca tgaaactgat
     5281 tgtcccggtg ctgttggatg cctccgtgcc accctatgac aagatccggg ttctgctgct
     5341 ctacatcctt ctgcggaacg gtgtgagtga ggaaaatctg gccaagctga tccagcatgc
     5401 taatgtgcag tcctacagca acctcatccg gaacctggag cagctggggg gcactgtcac
     5461 caactctgcg ggctcaggga cctctagccg gctggagcgg agagaacgca tggagcccac
     5521 ctaccagctg tctcgctggt ccccagtcat caaggacgtg atggaggatg tggtggagga
     5581 ccggctagac cgaaagctgt ggcccttcgt gtctgaccct gcccctgtgt ctagctccca
     5641 agcagctgtc agtgctcgct ttggccactg gcataaaaac aaagctggta tagaggctcg
     5701 ggctggaccc aggctcatcg tgtacattgt gggtggtgtc gccatgtctg aaatgagggc
     5761 tgcttatgag gtgaccagag ccactgaagg caagtgggag gtacttatag gctcttccca
     5821 catcctcact ccaacccgct tcctcgatga cctcaggacc ctggatcaga agctggagga
     5881 catagccctg ccctgagtcg acgagctcaa ctagtgcggc cgctttcgaa tctagagcct
     5941 gcagtctcga ggcatgcggt accaagcttg tcgagaagta ctagaggatc ataatcagcc
     6001 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     6061 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     6121 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6181 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg
     6241 agcctaggag atccgaacca gataagtgaa atctagttcc aaactatttt gtcattttta
     6301 attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta
     6361 aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac
     6421 agcggggcat ttttcttcct gttatgtttt taatcaaaca tcctgccaac tccatgtgac
     6481 aaaccgtcat cttcggctac tttttctctg tcacagaatg aaaatttttc tgtcatctct
     6541 tcgttattaa tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaatggcgaa
     6601 tgg
//
