LOCUS       sequence-225607-        5814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Rat synaptobrevin 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5814)
  AUTHORS   Lai Y, Choi UB, Leitz J, Rhee HJ, Lee C, Altas B, Zhao M, Pfuetzner 
            RA, Wang AL, Brose N, Rhee J, Brunger AT
  TITLE     Molecular Mechanisms of Synaptic Vesicle Priming by Munc13 and 
            Munc18.
  JOURNAL   Neuron. 2017 Aug 2;95(3):591-607.e10. doi: 
            10.1016/j.neuron.2017.07.004.
  PUBMED    28772123
REFERENCE   2  (bases 1 to 5814)
  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: 
            "10.1016/j.neuron.2017.07"; journalName: "Neuron"; date: "2017-08-2-
            2"; volume: "95"; issue: "3"; pages: "591-607.e10"
COMMENT     Sequence Label: pDC191
FEATURES             Location/Qualifiers
     source          1..5814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     117..136
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        117..135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    136..160
                     /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             175..197
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             219..236
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             258..278
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(651..669)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      665..712
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      749..1204
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(836..855)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1046..1067
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1230..1334
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1335..1403,1404..2195)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1553..1572)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     2936..2955
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3189..3206
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3221..3360
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3286..3305)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3446..3468
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3462..3653)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4462..5544)
                     /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     5506..5525
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5545..5622)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
       61 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
      121 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
      181 taactttaag aaggagatat acatatatgt cgtactacca tcaccatcac catcacgatt
      241 acgatatccc aactagtgaa aacctgtatt ttcagggcgc tagcgcgacg gcagccaccg
      301 tgccgcctgc agcccctgct ggtgaaggtg gcccaccggc accgccaccg aatctgactt
      361 gcaaccgtcg tctgcagcaa acgcaagcgc aggtcgacga ggttgttgat atcatgcgtg
      421 tcaatgttga caaagtcctg gagcgcgacc aaaagctgag cgaactggat gatcgtgcgg
      481 acgcgttgca ggcaggtgcc agccagttcg aaacctccgc tgcgaaattg aagcgcaaat
      541 actggtggaa gaacctgaaa atgatgatta ttctgggcgt gattagcgcg atcattctga
      601 tcattatcat cgtgtatttc agcacctaat cgagctcccg ggatccgcgg ccgctgagca
      661 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
      721 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
      781 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
      841 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
      901 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
      961 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1021 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1081 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1141 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1201 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1261 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1321 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1381 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1441 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1501 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1561 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1621 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1681 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1741 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1801 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1861 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     1921 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1981 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2041 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2101 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2161 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2221 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2281 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2341 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2401 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2461 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2521 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2581 tgctaatcct gttaccagtg gatcgacgga gcgatgccga gccgtgctga aacggtaaaa
     2641 ggttacgtgc aggccatgca ccagagctgg ttacaggatg tgctgctgcc agtggcgata
     2701 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2761 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2821 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2881 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2941 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3001 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3061 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3121 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3181 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3241 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     3301 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     3361 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3421 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3481 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     3541 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     3601 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     3661 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     3721 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     3781 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     3841 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     3901 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     3961 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     4021 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     4081 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     4141 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     4201 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     4261 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     4321 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     4381 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     4441 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4501 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     4561 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     4621 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     4681 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     4741 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     4801 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     4861 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     4921 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     4981 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     5041 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     5101 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     5161 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     5221 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     5281 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     5341 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     5401 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     5461 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     5521 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     5581 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     5641 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     5701 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     5761 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gccc
//
