LOCUS       sequence-369026-        6622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MUNC18-2 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6622)
  AUTHORS   Lai Y, Fois G, Flores JR, Tuvim MJ, Zhou Q, Yang K, Leitz J, Peters 
            J, Zhang Y, Pfuetzner RA, Esquivies L, Jones P, Frick M, Dickey BF, 
            Brunger AT
  TITLE     Inhibition of calcium-triggered secretion by hydrocarbon-stapled 
            peptides.
  JOURNAL   Nature. 2022 Mar;603(7903):949-956. doi: 10.1038/s41586-022-04543-1.
            Epub 2022 Mar 23.
  PUBMED    35322233
REFERENCE   2  (bases 1 to 6622)
  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"; doi: 
            "10.1038/s41586-022-04543-1"; journalName: "Nature"; date: 
            "2022-03"; volume: "603"; issue: "7903"; pages: "949-956"
COMMENT     Sequence Label: MUNC18-2
FEATURES             Location/Qualifiers
     source          1..6622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3605..3610,3611..3627,3628..3633))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3985..4076
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4001..4018
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4143..4160
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4182..4202
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    6212..6346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6249..6268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6303..6322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6375..6540
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     3481 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     3541 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     3601 ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca
     3661 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc tgtcctggct
     3721 ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg ccttgctgtt
     3781 cttctacggc aaggtgctgt gcacggatct gccctggctt caggagatcg gaagacctcg
     3841 gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca tcctcggttt
     3901 tctggaaggc gagcatcgtt tgttcgccca ggactctagc tatagttcta gtggttggct
     3961 acgtatactc cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa
     4021 ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc
     4081 cggattattc ataccgtccc accatcgggc gcggatctcg gtccgaaacc atgtcgtact
     4141 accatcacca tcaccatcac gattacgata tcccaacgac cgaaaacctg tattttcagg
     4201 gcgccatgga tccaatggcg cccttggggc tgaaggcggt ggtggtggaa aaaattctaa
     4261 gtggagtcat ccggagcgtt aagaaggatg gcgagtggaa ggtactcatc atggatcatc
     4321 cgagcatgcg aatcttgtca tcctgttgta aaatgtcaga tatcttggcg gagggcatca
     4381 ccattgtgga agacatcaac aaacggagag aacccattcc cagcttggag gcaatttatt
     4441 tgctgagtcc cacagagaag tcggttcagg ctctgattgc ggacttccag gggacaccaa
     4501 ccttcaccta caaagcagcc catatcttct tcacggacac gtgccccgag cccttgttca
     4561 gtgaactggg ccgctctcgc ctggcgaagg tggtgaagac actgaaagag atccacctcg
     4621 ccttcctccc ctatgaggcc caggtgttct ctctggatgc cccgcacagc acctataacc
     4681 tttattgccc atttcgggca ggggagcggg ggcggcagct cgatgtgctg gcccagcaga
     4741 tagccacact gtgtgccact cttcaggaat acccatccat tcgctaccgc aagggcccag
     4801 aagacacagc ccagctggcc catgctgtcc tggccaagct gaacgccttc aaggcagaca
     4861 ctcccagtct gggcgagggc ccagagaaaa cacgctcaca gttgctcata atggaccggg
     4921 cagtagaccc cgtgtccccg ctattacatg aactcacatt ccaagccatg gcctatgatc
     4981 ttctgcacat cgagcaggac acatacaggt atgagaccac aggactgagc gagtcccggg
     5041 agaaggctgt gctcctggat gaagacgatg acctgtgggt ggaactccgg cacatgcaca
     5101 tagcggatgt gtccaagaag gtcacggagc tcctgaagac attctgtgag agcaagcggc
     5161 tgaccacaga caaggccaac atcaaagacc tgtcccacat cctgaaaaag atgccacagt
     5221 atcagaagga gttgaacaag tactccacgc acctgcactt ggcagatgac tgcatgaagc
     5281 acttcaaggg ctcggtggag aagctgtgcg gtgtggagca ggacctggcc atgggctctg
     5341 atgcagaggg tgagaagatc aaggacacca tgaaactgat tgtcccggtg ctgttggatg
     5401 cctccgtgcc accctatgac aagatccggg ttctgctgct ctacatcctt ctgcggaacg
     5461 gtgtgagtga ggaaaatctg gccaagctga tccagcatgc taatgtgcag tcctacagca
     5521 acctcatccg gaacctggag cagctggggg gcactgtcac caactctgcg ggctcaggga
     5581 cctctagccg gctggagcgg agagaacgca tggagcccac ctaccagctg tctcgctggt
     5641 ccccagtcat caaggacgtg atggaggatg tggtggagga ccggctagac cgaaagctgt
     5701 ggcccttcgt gtctgaccct gcccctgtgt ctagctccca agcagctgtc agcagtgctc
     5761 gctttggcca ctggcataaa aacaaagctg gtatagaggc tcgggctgga cccaggctca
     5821 tcgtgtacat tgtgggtggt gtcgccatgt ctgaaatgag ggctgcttat gaggtgacca
     5881 gggccactga aggcaagtgg gaggtactta taggctcttc ccacatcctc actccaaccc
     5941 gcttcctcga tgacctcagg accctggatc agaagctgga ggacatagcc ctgccctgag
     6001 aattcaaagg cctacgtcga cgagctcaac tagtgcggcc gctttcgaat ctagagcctg
     6061 cagtctcgag gcatgcggta ccaagcttgt cgagaagtac tagaggatca taatcagcca
     6121 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     6181 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga
     6361 gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     6421 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     6481 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     6541 gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca
     6601 aaccgtcatc ttcggctact tt
//
