LOCUS       sequence-218717-        4928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rat Syntaxin 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4928)
  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 4928)
  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: pDC216
FEATURES             Location/Qualifiers
     source          1..4928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      578..1166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1067..1086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1320..1337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1435..1452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1436..1452
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1915..1934
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1915..1933
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1934..1958
                     /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             2013..2030
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2052..2072
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2947..2994
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3217..3236)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3242..3261)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3303..3394
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3395..3463,3464..4255)
                     /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     complement(3613..3632)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             join(4338..4928,1..492)
                     /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(4357..4376)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
       61 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
      121 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
      181 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
      241 gatagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
      301 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
      361 ctgttgccag tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gactcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact
      541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      721 ctgttcttct agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta
      781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      961 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1261 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1321 gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa
     1381 gcagaaggcc cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa
     1441 acgacggcca gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat
     1501 ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag
     1561 agcatttgtc agaatattta agggcgcctg tcactttgct tgatatatga gaattattta
     1621 accttataaa tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga
     1681 tgaacaccta taattaaact attcatctat tatttatgat tttttgtata tacaatattt
     1741 ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt
     1801 tttgatatca aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg
     1861 ttatcagtat ttattatcat ttagaataaa ttttgtgtcg cccttccgcg aaattaatac
     1921 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
     1981 acttttagga ggtaaaacat atgagctatt ggcaccatca ccaccaccac gactacgata
     2041 tcccgaccag cgagaatctg tattttcagg gtaaagatcg tacccaggag ctgcgtaccg
     2101 cgaaagacag cgacgatgat gacgacgtga ccgttacggt cgatcgtgac cgcttcatgg
     2161 acgaattctt cgagcaagtc gaagagatcc gcggctttat tgataagatc gcagagaacg
     2221 tggaagaggt taagcgcaaa cactccgcga ttctggcaag cccgaatcca gacgaaaaga
     2281 cgaaagaaga actggaagag ttgatgtccg acattaagaa aaccgcgaac aaagttcgta
     2341 gcaagctgaa atctatcgag cagagcatcg aacaggaaga gggtctgaac cgctccagcg
     2401 ccgatctgcg tattcgtaaa acgcagcaca gcactctgag ccgcaaattc gtcgaagtca
     2461 tgagcgagta caatgcaacc caaagcgact accgcgagcg cagcaaaggc cgtattcaac
     2521 gtcaactgga gatcaccggt cgtaccacca cctctgaaga gttggaagat atgctggaga
     2581 gcggtaaccc ggcgattttt gcgtctggca tcatcatgga cagcagcatc agcaagcaag
     2641 cgctgtgcga aattgaaacg cgccatagcg aaatcattaa actggagaac agcattcgtg
     2701 agttgcatga tatgttcatg gacatggcaa tgctggttga gagccagggt gagatgatcg
     2761 atcgtattga atacaatgtg gagcatgccg ttgattatgt cgagcgtgcg gtgagcgaca
     2821 cgaagaaggc cgttaagtac cagtcgaaag ctcgtcgtaa gaagatcatg attatcatct
     2881 ctagcgtgat tctgggtatt atcattgctt ccacgatcgg tggcattttt ggctaactcg
     2941 agccccctag cataacccct tggggcctct aaacgggtct tgaggggttt tttgcccctg
     3001 agacgcgtca atcgagttcg tacctaaggg cgacaccccc taattagccc gggcgaaagg
     3061 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
     3121 tggggagtcc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
     3181 tcaggtggga ccaccgcgct actgccgcca ggcaaacaag gggtgttatg agccatattc
     3241 aggtataaat gggctcgcga taatgttcag aattggttaa ttggttgtaa cactgacccc
     3301 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3361 ataaatgctt caataatatt gaaaaaggaa gaatatgagt attcaacatt tccgtgtcgc
     3421 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3481 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3541 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3601 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3661 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3721 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3781 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3841 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3901 agccatacca aacgacgagc gtgacaccac gatgcctgta gcgatggcaa caacgttgcg
     3961 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4021 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4081 tgctgataaa tccggagccg gtgagcgtgg ttctcgcggt atcatcgcag cgctggggcc
     4141 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4201 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaagcggc
     4261 gcgccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     4321 aattcagggt ggtgaatatg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     4381 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     4441 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     4501 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     4561 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     4621 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     4681 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     4741 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     4801 ccatcaacag tattattttc tcccatgagg acggtacgcg actgggcgtg gagcatctgg
     4861 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     4921 gtctgcgt
//
