LOCUS       sequence-204177-        5218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Complexin-1 fragment (1-83) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5218)
  AUTHORS   Zhou Q, Zhou P, Wang AL, Wu D, Zhao M, Sudhof TC, Brunger AT
  TITLE     The primed SNARE-complexin-synaptotagmin complex for neuronal 
            exocytosis.
  JOURNAL   Nature. 2017 Aug 24;548(7668):420-425. doi: 10.1038/nature23484. 
            Epub 2017 Aug 16.
  PUBMED    28813412
REFERENCE   2  (bases 1 to 5218)
  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/nature23484"; journalName: "Nature"; date: "2017-08-24- 
            24"; volume: "548"; issue: "7668"; pages: "420-425"
COMMENT     Sequence Label: pQZ083
FEATURES             Location/Qualifiers
     source          1..5218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      31..619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     520..539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     773..790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        863..940
                     /gene="lacI"
                     /label=lacI promoter
     CDS             941..2023
                     /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(960..979)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2146..2673
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2166..2183)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2166..2182)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2175..2197)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2775..2780,2781..2796,2797..2803)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2811..2827
                     /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             2850..3503
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3461..3483
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3510..3533
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3860..3882)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4027..4045)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4113..4217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4218..4286,4287..5078)
                     /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"
     primer_bind     complement(4436..4455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
       61 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
      181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      661 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      721 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      781 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
      841 tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc
      901 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
      961 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     1021 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     1081 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     1141 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     1201 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     1261 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     1321 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     1381 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     1441 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     1501 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     1561 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     1621 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     1681 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     1741 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     1801 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     1861 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     1921 aaccacgcta gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2221 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2281 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
     2341 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
     2401 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
     2461 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
     2521 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
     2581 ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     2641 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     2701 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     2761 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
     2821 ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat tggaaaatta
     2881 agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa tatgaagagc
     2941 atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg
     3001 agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag tctatggcca
     3061 tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag
     3121 agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg agaattgcat
     3181 atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct gaaatgctga
     3241 aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat gtaacccatc
     3301 ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca atgtgcctgg
     3361 atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca caaattgata
     3421 agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa gccacgtttg
     3481 gtggtggcga ccatcctcca aaatcggatc tggaagttct gttccagggg cccctgggat
     3541 ccatggagtt cgtgatgaaa caagccctgg gaggggccac caaggatatg gggaagatgc
     3601 ttgggggtga tgaggagaag gaccccgatg ccgccaagaa ggaggaggag cggcaggagg
     3661 cactgcggca ggcagaggag gagcgcaaag caaagtatgc caagatggag gcagaacgtg
     3721 aggtcatgcg gcagggcata cgagataagt atggcatcaa gaagaaggag gagcgtgagg
     3781 ctgaggctca gtaactcgag cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt
     3841 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3901 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3961 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac
     4021 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4081 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4141 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4201 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4261 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4321 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4381 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4441 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     4501 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4561 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4621 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4681 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4741 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4801 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4861 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4921 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4981 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5041 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5101 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5161 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtc
//
