LOCUS       sequence-128806-        6683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses Syt1-SNARE37aa-linker complex with pDC184.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6683)
  AUTHORS   Zhou Q, Lai Y, Bacaj T, Zhao M, Lyubimov AY, Uervirojnangkoorn M, 
            Zeldin OB, Brewster AS, Sauter NK, Cohen AE, Soltis SM, Alonso-Mori 
            R, Chollet M, Lemke HT, Pfuetzner RA, Choi UB, Weis WI, Diao J, 
            Sudhof TC, Brunger AT
  TITLE     Architecture of the synaptotagmin-SNARE machinery for neuronal 
            exocytosis.
  JOURNAL   Nature. 2015 Sep 3;525(7567):62-7. doi: 10.1038/nature14975. Epub 
            2015 Aug 17.
  PUBMED    26280336
REFERENCE   2  (bases 1 to 6683)
  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/nature14975"; journalName: "Nature"; date: "2015-09-3- 3"; 
            volume: "525"; issue: "7567"; pages: "62-7"
COMMENT     Sequence Label: p045
FEATURES             Location/Qualifiers
     source          1..6683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        1297..1315
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1316..1340
                     /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             1629..1673
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1711..1729)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1725..1772
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1809..2264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1896..1915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2106..2127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2379..3170,3171..3239))
                     /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     3002..3021
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3240..3332)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3413..4001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3902..3921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4155..4172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4187..4329
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4255..4274)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4415..4437
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4431..4622)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5194..6276)
                     /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     6238..6257
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6277..6354)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6560..6579
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(6667..6683,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atggagaaac tgggaaagct ccaatattca ctggactatg acttccagaa
      121 taaccagctg ttggtgggaa tcatccaggc tgctgaactg cccgccctgg acatgggggg
      181 tacatccgat ccatacgtca aagtcttcct gctgcctgac aaaaagaaga aatttgagac
      241 taaagtccac cggaaaaccc tcaatccagt cttcaatgaa caatttactt tcaaggtacc
      301 ctactcggaa ttaggtggca aaaccctggt gatggctgtg tatgactttg atcgcttctc
      361 caagcacgac atcatcggag agttcaaagt tcctatgaac accgtggatt ttggccatgt
      421 gaccgaggag tggcgcgatc tccagagcgc tgagaaagaa gagcaagaga aactgggtga
      481 catctgcttc tccctccgct acgtccctac tgccggcaaa ctgactgttg tcattctgga
      541 agccaagaac ctgaagaaga tggatgtggg tggcttatct gatccctacg tgaagattca
      601 cctgatgcag aacggtaaga ggctgaagaa gaaaaagacg acgattaaga agaacacact
      661 caacccctac tacaacgagt ccttcagctt tgaagttccg ttcgagcaaa tccagaaagt
      721 gcaagtggtg gtaactgttt tggactatga caagattggc aagaacgacg ccatcggcaa
      781 agtcttcgtt ggttacaaca gcactggggc ggagctgcga cactggtcag acatgctggc
      841 caacccccgg cgacccatcg cacagtggca cactctgcag gtagaggagg aggttgatgc
      901 catgctggct gtcaagaaga acctgtcgtc ggatagctcc ttgtcctctc cgtccgcttt
      961 gaatagcctg tcctctcctt cggccctgaa ctcgacggca agcaatagtc cgggaattga
     1021 ggggctgagc gcccgggaaa atgagatgga tgagaacctg gagcaggtga gcggcatcat
     1081 cggaaacctc cgccatatgg cactagacat gggcaatgag attgacaccc agaatcgcca
     1141 gatcgacagg atcatggaga aggctgattc caacaaaacc agaattgatg aagccaacca
     1201 acgtgcaaca aagatgctgg gctaaaagct tgcggccgca taatgcttaa gtcgaacaga
     1261 aagtaatcgt attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat
     1321 tgtgagcgga taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac
     1381 atatgcgcaa tgagctggag gagatgcaga ggagggctga ccagctggct gatgagtccc
     1441 tggaaagcac ccgtcgcatg ctgcagctgg ttgaagagag taaagatgct ggcatcagga
     1501 ctttggttat gttggatgag caaggcgaac aactcgatcg tgtcgaagaa ggcatgaacc
     1561 atatcaacca agacatgaag gaggccgaga aaaatttaaa agatttaggc aaataactcg
     1621 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     1681 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     1741 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     1801 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1861 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1921 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1981 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2041 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2101 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2161 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2221 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttctggcg gcacgatggc
     2281 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2341 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2401 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2461 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2521 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2581 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2641 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2701 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2761 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2821 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2881 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2941 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3001 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3061 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3121 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3181 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3241 ctcttccttt ttcaatcatg attgaagcat ttatcagggt tattgtctca tgagcggata
     3301 catatttgaa tgtatttaga aaaataaaca aataggtcat gaccaaaatc ccttaacgtg
     3361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3541 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4081 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4141 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4201 ttctccttac gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat
     4261 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     4321 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4381 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4441 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4501 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4561 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4621 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4681 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4741 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4801 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4861 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4921 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4981 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     5041 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac
     5101 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     5161 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5221 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5281 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     5341 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     5401 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     5461 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     5521 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     5581 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     5641 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     5701 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     5761 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     5821 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     5881 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     5941 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     6001 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     6061 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     6121 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     6181 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     6241 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     6301 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     6361 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     6421 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     6481 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     6541 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     6601 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     6661 cgaaattaat acgactcact ata
//
