LOCUS       sequence-204169-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses trans SNARE complex with pQZ210 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5729)
  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 5729)
  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: pDC183
FEATURES             Location/Qualifiers
     source          1..5729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    10..34
                     /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             49..71
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        353..371
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    372..396
                     /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             685..729
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(767..785)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      781..828
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      865..1320
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(952..971)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1162..1183
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1435..2226,2227..2295))
                     /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     2058..2077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2296..2388)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2469..3057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2958..2977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3211..3228
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3243..3382
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3308..3327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3468..3490
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3484..3675)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4247..5329)
                     /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     5291..5310
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5330..5407)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5613..5632
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(5720..5729,1..9)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
       61 taagaaggag atataccatg gcccgggaaa atgagatgga tgagaacctg gagcaggtga
      121 gcggcatcat cggaaacctc cgccatatgg cactagacat gggcaatgag attgacaccc
      181 agaatcgcca gatcgacagg atcatggaga aggctgattc caacaaaacc agaattgatg
      241 aagccaacca acgtgcaaca aagatgctgg gctaagtcga caagcttgcg gccgcataat
      301 gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga
      361 ctcactatag gggaattgtg agcggataac aattccccat cttagtatat tagttaagta
      421 taagaaggag atatacatat gcgcaatgag ctggaggaga tgcagaggag ggctgaccag
      481 ctggctgatg agtccctgga aagcacccgt cgcatgctgc agctggttga agagagtaaa
      541 gatgctggca tcaggacttt ggttatgttg gatgagcaag gcgaacaact cgatcgtgtc
      601 gaagaaggca tgaaccatat caaccaagac atgaaggagg ccgagaaaaa tttaaaagat
      661 ttaggcaaat aactcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac
      721 atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc tgagcaataa
      781 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
      841 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
      901 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
      961 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1021 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1081 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1141 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1201 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1261 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     1321 ctggcggcac gatggcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1381 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1441 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1501 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1561 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1621 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1681 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1741 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1801 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1861 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1921 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1981 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2041 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2101 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2161 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2221 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2281 atgttgaata ctcatactct tcctttttca atcatgattg aagcatttat cagggttatt
     2341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggtcatgacc
     2401 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2461 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2521 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2581 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2641 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2701 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2761 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2821 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2881 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2941 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3001 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3061 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3121 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3181 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3241 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     3301 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     3361 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3421 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3481 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     3541 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     3601 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     3661 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     3721 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3781 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3841 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3901 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3961 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     4021 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     4081 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat
     4141 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     4201 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     4261 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4321 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     4381 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     4441 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     4501 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
     4561 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     4621 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     4681 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     4741 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     4801 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     4861 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     4921 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     4981 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     5041 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     5101 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     5161 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     5221 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     5281 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     5341 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     5401 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     5461 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     5521 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     5581 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     5641 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     5701 gatctcgatc ccgcgaaatt aatacgact
//
