LOCUS       sequence-387507-        6212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Synaptotagmin 2 in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6212)
  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 6212)
  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: Syt2
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(103..122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      916..1504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1405..1424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1658..1675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1748..1825
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1826..2908
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1845..1864)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2957..2962,2963..2980,2981..2987)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2995..3011
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3019..3035
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3019..3035
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3031..3558
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(3051..3068)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3051..3067)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3060..3082)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(3660..3665,3666..3681,3682..3688)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3696..3712
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             3735..4388
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4346..4368
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4395..4412
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(5739..5761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5906..5924)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5992..6096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6097..6165,6166..6212,1..745)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
       61 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      121 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      181 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      241 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      301 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      361 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      421 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      481 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      541 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      601 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      661 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      721 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
      781 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1021 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1621 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1681 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     1741 aaattccgac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
     1801 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     1861 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     1921 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     1981 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     2041 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     2101 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     2161 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     2221 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     2281 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     2341 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     2401 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     2461 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     2521 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     2581 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     2641 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     2701 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     2761 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     2821 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2881 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
     2941 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     3001 gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc
     3061 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     3121 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3181 cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg
     3241 ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac
     3301 tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc cattacggtc
     3361 aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt
     3421 gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt tggaattagc
     3481 ttatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta
     3541 tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct
     3601 ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt
     3661 tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac
     3721 aggaaacagt attcatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac
     3781 ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg
     3841 atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc
     3901 cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag
     3961 ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg
     4021 aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg
     4081 aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc
     4141 gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt
     4201 atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat
     4261 tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca
     4321 gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc
     4381 ctccaaaatc ggatctggtt ccgcgtggat ccatgagaaa catcttcaag aggaaccagg
     4441 agcccattgt ggctccggcc accaccactg ccacaatgcc tctggcaccc gccgcacctg
     4501 ccgataactc tacagagagc acgggcaccg gggagagcca agaagacatg ttcgccaagc
     4561 tgaaggacaa attcttcaat gagatcaaca agatcccttt gcccccctgg gctctgattg
     4621 ccatggccgt ggttgctggc ctcctgctgc tcacctgctg cttctgcatc tgtaagaagt
     4681 gctgctgcaa gaagaagaaa aacaagaagg agaagggcaa aggcatgaag aacgccatga
     4741 acatgaagga catgaagggg ggccaggatg atgacgacgc ggagacaggc ctgactgaag
     4801 gagaaggaga aggcgaggag gagaaagagc cggagaacct gggcaaattg cagttttctc
     4861 tggactatga tttccaagcc aaccagctca ccgtgggcgt cctgcaggct gctgaactcc
     4921 cggccctgga catgggtggc acgtcagacc cttacgtcaa agtcttcctc ctcccagaca
     4981 agaagaagaa atatgagacc aaggtgcacc ggaagacact gaacccagcc ttcaacgaaa
     5041 ctttcacttt caaggtgcca taccaggagt taggaggcaa aaccctggtg atggctatct
     5101 atgactttga ccgcttctct aagcatgaca tcatcgggga ggtgaaagtg cccatgaaca
     5161 cggtggacct tggccagccc atcgaggaat ggagagacct acaaggcgga gagaaggaag
     5221 agccagagaa gctgggtgac atctgtacct ccttgcgcta cgtgcccact gctgggaagc
     5281 tcaccgtctg tatcctggag gccaagaacc tgaagaagat ggatgtgggg ggcctctcag
     5341 acccctatgt gaagatccac ttgatgcaga atggcaagag actcaagaag aagaagacga
     5401 cggtgaagaa gaagaccttg aacccctact tcaatgagtc attcagcttc gagatcccct
     5461 ttgagcagat ccagaaagtc caggtggtcg tcaccgtgct agactatgac aaactgggca
     5521 agaatgaagc catcggaaag atcttcgtag gcagcaacgc tacaggcacg gagctgcggc
     5581 actggtccga catgctggcg aaccctcgga ggcccatcgc ccagtggcac tctctgaagc
     5641 ctgaggaaga agtggatgct cttctgggca agaacaagta gaattcatcg tgactgactg
     5701 acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5761 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5821 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     5881 gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5941 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6001 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6061 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6121 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6181 gtgaaagtaa aagatgctga agatcagttg gg
//
