LOCUS       sequence-384261-        5799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid encoding GST-tagged SYT9 C2AB domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5799)
  AUTHORS   Seibert MJ, Evans CS, Stanley KS, Wu Z, Chapman ER
  TITLE     Synaptotagmin 9 Modulates Spontaneous Neurotransmitter Release in 
            Striatal Neurons by Regulating Substance P Secretion.
  JOURNAL   J Neurosci. 2023 Mar 1;43(9):1475-1491. doi: 
            10.1523/JNEUROSCI.1857-22.2023. Epub 2023 Feb 2.
  PUBMED    36732068
REFERENCE   2  (bases 1 to 5799)
  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: 
            "10.1523/JNEUROSCI.1857-22.2023"; journalName: "J Neurosci"; date: 
            "2023-03-1- 1"; volume: "43"; issue: "9"; pages: "1475-1491"
COMMENT     Sequence Label: pGEX SYT9 C2AB
FEATURES             Location/Qualifiers
     source          1..5799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(154..159,160..175,176..182)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    190..206
                     /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             229..882
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     840..862
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             889..906
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1820..1842)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1987..2005)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2073..2177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2178..2246,2247..3038)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2396..2415)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3209..3797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3698..3717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3951..3968
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4041..4118
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4119..5201
                     /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(4138..4157)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5250..5255,5256..5273,5274..5280)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5288..5304
                     /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)."
     primer_bind     5312..5328
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5312..5328
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5324..5799,1..52)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5344..5361)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5344..5360)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5353..5375)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
       61 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
      121 taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa
      181 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat gtcccctata
      241 ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt
      301 gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa
      361 aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa
      421 ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt
      481 tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac
      541 ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc
      601 aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat
      661 ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac
      721 atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa
      781 gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag
      841 ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct ggttccgcgt
      901 ggatccatgt cagacaaaca ccagctaggc cgactgcagt attcactgga ctatgacttc
      961 cagactggcc agctcctggt gggcatcttg caagctcagg gactggctgc gttggaccta
     1021 ggaggttcct cagatcccta tgttagtgtc tatctgctgc cagacaagcg gaggcgacat
     1081 gagaccaagg tgcatcggca gaccctgaat ccacactttg gagagacatt tgccttcaag
     1141 gtcccatacg tggaactagg gggcagagtg ctggtcatgg cggtgtatga ctttgatcgg
     1201 ttctcccgca acgatgccat cggggaggtg cgggtgccta tgagttcagt gaacctgggg
     1261 cggccggtgc aggcctggag agagctgcag gtggctccca aagaggagca agagaaactg
     1321 ggggacatct gcttctctct ccgttatgtc cccacggccg ggaagctcac cgtcattgtc
     1381 ttggaagcta aaaacctgaa gaaaatggat gtaggaggac tctcagatcc ctatgtgaag
     1441 gtgcacctgc tccagggagg taaaaaggtt cgcaagaaga aaaccaccat taagaagaac
     1501 accctgaacc cctattacaa cgaggccttc agctttgagg tgccctgtga ccaggtgcag
     1561 aaagtccagg tggagctgac tgttctggat tatgacaaac tggggaagaa tgaggccatc
     1621 gggagagtgg cggtgggcgc agcagtcggt ggggctggcc tgcggcactg ggctgacatg
     1681 ttggccaacc ccaggcgccc cattgcccag tggcactctc tgcggccccc tgacagagcc
     1741 aggccaatac ctgcaccctg agaattcatc gtgactgact gacgatctgc ctcgcgcgtt
     1801 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     1861 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     1921 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac
     1981 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2041 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2101 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2161 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2221 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2281 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2341 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2401 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2461 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2521 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2581 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2641 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2701 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2761 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2821 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2881 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2941 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3001 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3061 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3121 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3181 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3241 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3301 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3361 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3421 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3481 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3541 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3601 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3661 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3721 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3781 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3841 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3901 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3961 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4021 tttcacaccg cataaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
     4081 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     4141 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     4201 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     4261 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     4321 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     4381 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     4441 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     4501 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     4561 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     4621 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     4681 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     4741 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     4801 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     4861 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     4921 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     4981 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     5041 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     5101 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5161 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
     5221 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     5281 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga
     5341 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5401 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5461 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc
     5521 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
     5581 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta
     5641 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
     5701 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg
     5761 cgttggaatt agcttatcga ctgcacggtg caccaatgc
//
