LOCUS       sequence-383657-        5346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid encoding GST-tagged SYT9 C2A domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5346)
  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 5346)
  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 C2A
FEATURES             Location/Qualifiers
     source          1..5346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(3..8,9..24,25..31)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    39..55
                     /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             78..731
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     689..711
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             738..755
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1216..1238)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1383..1401)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1469..1573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1574..1642,1643..2434)
                     /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(1792..1811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2605..3193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3094..3113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3347..3364
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3437..3514
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3515..4597
                     /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(3534..3553)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4646..4651,4652..4669,4670..4676)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4684..4700
                     /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     4708..4724
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4708..4724
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4720..5247
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4740..4757)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4740..4756)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4749..4771)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      121 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      181 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      241 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      301 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      361 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      421 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      481 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      541 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      601 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      661 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      721 atcctccaaa atcggatctg gttccgcgtg gatccatgtc agacaaacac cagctaggcc
      781 gactgcagta ttcactggac tatgacttcc agactggcca gctcctggtg ggcatcttgc
      841 aagctcaggg actggctgcg ttggacctag gaggttcctc agatccctat gttagtgtct
      901 atctgctgcc agacaagcgg aggcgacatg agaccaaggt gcatcggcag accctgaatc
      961 cacactttgg agagacattt gccttcaagg tcccatacgt ggaactaggg ggcagagtgc
     1021 tggtcatggc ggtgtatgac tttgatcggt tctcccgcaa cgatgccatc ggggaggtgc
     1081 gggtgcctat gagttcagtg aacctggggc ggccggtgca ggcctggaga gagctgcagg
     1141 tggctcccaa agagtgagaa ttcatcgtga ctgactgacg atctgcctcg cgcgtttcgg
     1201 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1261 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1321 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa
     1381 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1441 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1501 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1561 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1621 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1681 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1741 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1801 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     1861 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1921 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1981 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2041 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2101 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2161 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2221 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2281 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2341 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2401 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2461 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2521 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2581 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2641 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2701 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2761 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2821 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2881 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2941 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3001 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3061 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3121 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3181 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3241 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3301 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3361 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3421 acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     3481 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     3541 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     3601 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     3661 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3721 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     3781 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     3841 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     3901 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     3961 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     4021 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     4081 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     4141 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     4201 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     4261 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     4321 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     4381 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     4441 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     4501 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4561 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     4621 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4681 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca
     4741 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4801 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4861 ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca
     4921 gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc
     4981 ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc
     5041 attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca
     5101 tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt
     5161 ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
     5221 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
     5281 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
     5341 atgagc
//
