LOCUS       sequence-368913-        4927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Syntaxin 3 in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4927)
  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 4927)
  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: Stx3
FEATURES             Location/Qualifiers
     source          1..4927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      239..827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     728..747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     981..998
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1096..1113
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1097..1113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1576..1595
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1576..1594
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1595..1619
                     /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)."
     RBS             1647..1655
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1664..1681
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1682..1702
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2312..2323
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      2607..2654
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2877..2896)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2902..2921)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2963..3054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3055..3123,3124..3915)
                     /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(3273..3292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             join(3998..4927,1..153)
                     /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(4017..4036)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 caggcggtga agggcaatca gctgttgcca gtctcactgg tgaaaagaaa aaccaccctg
       61 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      121 cgacaggttt cccgactgga aagcgggcag tgactcatga ccaaaatccc ttaacgtgag
      181 ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      361 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttagcc caccacttca
      421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      721 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      781 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      841 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      901 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      961 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaggc gagagtaggg
     1021 aactgccagg catcaaacta agcagaaggc ccctgacgga tggccttttt gcgtttctac
     1081 aaactctttc tgtgttgtaa aacgacggcc agtcttaagc tcgggccccc tgggcggttc
     1141 tgataacgag taatcgttaa tccgcaaata acgtaaaaac ccgcttcggc gggttttttt
     1201 atggggggag tttagggaaa gagcatttgt cagaatattt aagggcgcct gtcactttgc
     1261 ttgatatatg agaattattt aaccttataa atgagaaaaa agcaacgcac tttaaataag
     1321 atacgttgct ttttcgattg atgaacacct ataattaaac tattcatcta ttatttatga
     1381 ttttttgtat atacaatatt tctagtttgt taaagagaat taagaaaata aatctcgaaa
     1441 ataataaagg gaaaatcagt ttttgatatc aaaattatac atgtcaacga taatacaaaa
     1501 tataatacaa actataagat gttatcagta tttattatca tttagaataa attttgtgtc
     1561 gcccttccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     1621 ctctagaaat aattttgttt aacttttaag gaggtaaaaa atgcatcatc atcatcacca
     1681 cgaaaatctg tatttccaat ctggcattag cggtggcggc ggcggcattc tgatgaaaga
     1741 tcgtctggag caactgaaag cgaaacagct gacgcaagac gacgacaccg acgctgttga
     1801 gatcgcgatc gacaacactg cgttcatgga cgaattcttt tctgagatcg aagagactcg
     1861 cctgaatatt gataagatca gcgaacacgt ggaagaggcg aaaaagttgt atagcatcat
     1921 cctgtccgcc ccgatcccgg agccgaaaac caaagacgac ctggagcaat tgacgaccga
     1981 gatcaaaaag cgtgcaaaca acgttcgcaa caagctgaag tccatggaaa agcatattga
     2041 agaagatgaa gtccgttcca gcgcggatct gcgtattcgt aagagccagc acagcgtcct
     2101 gagccgtaag tttgtcgaag tgatgaccaa gtacaacgaa gcccaagtgg actttcgcga
     2161 gcgcagcaag ggtcgtatcc agcgtcagct ggaaattacg ggtaaaaaga ccaccgatga
     2221 agaattagaa gaaatgctgg agtcgggtaa tccggccatt tttacgagcg gtatcattga
     2281 cagccagatc agcaaacaag cactgagcga gattgagggt cgccacaaag atattgttcg
     2341 tctggagagc agcattaaag aattgcacga tatgttcatg gatatcgcga tgctggtcga
     2401 gaatcagggc gagatgctcg ataatattga gctgaacgtt atgcacaccg tcgaccacgt
     2461 cgagaaagcg cgttgcgaaa ccaaaaaagc tgtgaagtac cagagccagg cacgcaagaa
     2521 actgattatc attatcgtgt tggttgtggt gctgctgggt atccttgcac tgatcattgg
     2581 tctgagcgtt ggtctgaatt aaccccctag cataacccct tggggcctct aaacgggtct
     2641 tgaggggttt tttgcccctg agacgcgtca atcgagttcg tacctaaggg cgacacccca
     2701 taattagccc gggcgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     2761 ggcagttccc tactctcgca tggggagtcc ccacactacc atcggcgcta cggcgtttca
     2821 cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaacaag
     2881 gggtgttatg agccatattc aggtataaat gggctcgcga taatgttcag aattggttaa
     2941 ttggttgtaa cactgacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3001 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gaatatgagt
     3061 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3121 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3181 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3241 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3301 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3361 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3421 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3481 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3541 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3601 gcgatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3661 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3721 cttccggctg gctggtttat tgctgataaa tccggagccg gtgagcgtgg ttctcgcggt
     3781 atcatcgcag cgctggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3841 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3901 attaagcatt ggtaagcggc gcgccatcga atggcgcaaa acctttcgcg gtatggcatg
     3961 atagcgcccg gaagagagtc aattcagggt ggtgaatatg aaaccagtaa cgttatacga
     4021 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     4081 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     4141 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     4201 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     4261 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     4321 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     4381 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     4441 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgagg acggtacgcg
     4501 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     4561 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     4621 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     4681 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     4741 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     4801 ctcggtagtg ggatacgacg ataccgaaga tagctcatgt tatatcccgc cgttaaccac
     4861 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     4921 tcagggc
//
