LOCUS       sequence-383650-        5469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid encoding GST-tagged SYT1 C2B domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5469)
  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 5469)
  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 SYT1 C2B
FEATURES             Location/Qualifiers
     source          1..5469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(70..75,76..91,92..98)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    106..122
                     /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             145..798
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     756..778
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             805..822
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1406..1428)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1573..1591)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1659..1763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1764..1832,1833..2624)
                     /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(1982..2001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2795..3383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3284..3303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3537..3554
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3627..3704
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3705..4787
                     /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(3724..3743)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4836..4841,4842..4859,4860..4866)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4874..4890
                     /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     4898..4914
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4898..4914
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4910..5437
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4930..4947)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4930..4946)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4939..4961)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
       61 aatgagctgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac
      121 aatttcacac aggaaacagt attcatgtcc cctatactag gttattggaa aattaagggc
      181 cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg
      241 tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt
      301 cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata
      361 cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt
      421 tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt
      481 aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg
      541 ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac
      601 ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg
      661 ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac
      721 ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt
      781 ggcgaccatc ctccaaaatc ggatctggtt ccgcgtggat ccatgaccgt ggattttggc
      841 catgtgaccg aggagtggcg cgatctccag agcgctgaga aagaagagca agagaaactg
      901 ggtgacatct gcttctccct ccgctacgtc cctactgccg gcaaactgac tgttgtcatt
      961 ctggaagcca agaacctgaa gaagatggat gtgggtggct tatctgatcc ctacgtgaag
     1021 attcacctga tgcagaacgg taagaggctg aagaagaaaa agacgacgat taagaagaac
     1081 acactcaacc cctactacaa cgagtccttc agctttgaag ttccgttcga gcaaatccag
     1141 aaagtgcaag tggtggtaac tgttttggac tatgacaaga ttggcaagaa cgacgccatc
     1201 ggcaaagtct tcgttggtta caacagcact ggggcggagc tgcgacactg gtcagacatg
     1261 ctggccaacc cccggcgacc catcgcacag tggcacactc tgcaggtaga ggaggaggtt
     1321 gatgccatgc tggctgtcaa gaagtaagaa ttcatcgtga ctgactgacg atctgcctcg
     1381 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     1441 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     1501 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt
     1561 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1621 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1681 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1741 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1801 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1861 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1921 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1981 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2041 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2101 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2161 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2221 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2281 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     2341 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2401 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2461 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2521 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2581 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2641 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2701 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2941 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3481 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3541 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3601 gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta
     3661 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     3721 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     3781 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     3841 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     3901 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     3961 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     4021 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     4081 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     4141 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     4201 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     4261 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     4321 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     4381 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     4441 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     4501 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     4561 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     4621 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     4681 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4741 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4801 aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     4861 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     4921 tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4981 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5041 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt
     5101 tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac
     5161 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     5221 gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     5281 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     5341 tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
     5401 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     5461 caaggcgca
//
