LOCUS       sequence-224901-        6097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Rat SNAP25.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6097)
  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 6097)
  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: pDC158
FEATURES             Location/Qualifiers
     source          1..6097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..37
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     121..140
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        121..139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    140..164
                     /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             179..201
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             223..240
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             262..282
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(938..956)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      952..999
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1036..1491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1123..1142)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1333..1354
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1517..1621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1622..1690,1691..2482)
                     /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(1840..1859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3223..3242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3476..3493
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3508..3647
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3573..3592)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3733..3755
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3749..3940)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4749..5831)
                     /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     5793..5812
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5832..5909)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
       61 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
      121 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
      181 tgtttaactt taagaaggag atatacatat atgtcgtact accatcacca tcaccatcac
      241 gattacgata tcccaactag tgaaaacctg tattttcagg gcgctagcat ggccgaagac
      301 gcggatatgc gcaatgagct ggaggagatg cagaggaggg ctgaccagct ggctgatgag
      361 tccctggaaa gcacccgtcg catgctgcag ctggttgaag agagtaaaga tgctggcatc
      421 aggactttgg ttatgttgga tgagcaaggc gaacaactcg atcgtgtcga agaaggcatg
      481 aaccatatca accaagacat gaaggaggcc gagaaaaatt taaaagattt aggcaaatcc
      541 tctggccttt tcatatctcc ttctaacaag cttaaatcca gtgatgctta caaaaaagcc
      601 tggggcaata atcaggatgg agtagtggcc agccagcccg cccgtgtggt ggatgaacgg
      661 gagcagatgg ccatcagtgg tggcttcatc cgcagggtaa caaatgatgc ccgggaaaat
      721 gagatggatg agaacctgga gcaggtgagc ggcatcatcg gaaacctccg ccatatggca
      781 ctagacatgg gcaatgagat tgacacccag aatcgccaga tcgacaggat catggagaag
      841 gctgattcca acaaaaccag aattgatgaa gccaaccaac gtgcaacaaa gatgctgggc
      901 agtgggtaat agtgactcga gctcccggga tccgcggccg ctgagcaata actagcataa
      961 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1021 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1081 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1141 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1201 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1261 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1321 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1381 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1441 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1501 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1561 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1621 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1681 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1741 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1801 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1861 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1921 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1981 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2041 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2101 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2161 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2221 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2281 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2341 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2401 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2461 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2521 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2581 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2761 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2881 accagtggat cgacggagcg atgccgagcc gtgctgaaac ggtaaaaggt tacgtgcagg
     2941 ccatgcacca gagctggtta caggatgtgc tgctgccagt ggcgataagt cgtgtcttac
     3001 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3061 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3121 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3181 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3241 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3301 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3361 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3421 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3481 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     3541 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3601 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3661 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3721 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3781 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     3841 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3901 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     3961 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4021 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4081 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     4141 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     4201 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     4261 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     4321 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     4381 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
     4441 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     4501 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     4561 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4621 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4681 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     4741 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4801 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4861 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4921 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4981 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     5041 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     5101 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     5161 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     5221 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5281 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     5341 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     5401 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     5461 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     5521 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     5581 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5641 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5701 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5761 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5821 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5881 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     5941 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     6001 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     6061 cacgccgaaa caagcgctca tgagcccgaa gtggcga
//
