LOCUS       sequence-384264-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid encoding GST-tagged SYT9 C2B domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  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 5406)
  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 C2B
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(69..74,75..90,91..97)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    105..121
                     /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             144..797
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     755..777
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             804..821
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1342..1364)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1509..1527)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1595..1699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1700..1768,1769..2560)
                     /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(1918..1937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2731..3319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3220..3239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3473..3490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3563..3640
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3641..4723
                     /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(3660..3679)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4772..4777,4778..4795,4796..4802)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4810..4826
                     /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     4834..4850
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4834..4850
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4846..5373
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4866..4883)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4866..4882)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4875..4897)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
       61 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
      121 atttcacaca ggaaacagta ttcatgtccc ctatactagg ttattggaaa attaagggcc
      181 ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt
      241 atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc
      301 ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac
      361 gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt
      421 caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta
      481 aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt
      541 tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact
      601 tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc ctggatgcgt
      661 tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt gataagtact
      721 tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg tttggtggtg
      781 gcgaccatcc tccaaaatcg gatctggttc cgcgtggatc caaagaggag caagagaaac
      841 tgggggacat ctgcttctct ctccgttatg tccccacggc cgggaagctc accgtcattg
      901 tcttggaagc taaaaacctg aagaaaatgg atgtaggagg actctcagat ccctatgtga
      961 aggtgcacct gctccaggga ggtaaaaagg ttcgcaagaa gaaaaccacc attaagaaga
     1021 acaccctgaa cccctattac aacgaggcct tcagctttga ggtgccctgt gaccaggtgc
     1081 agaaagtcca ggtggagctg actgttctgg attatgacaa actggggaag aatgaggcca
     1141 tcgggagagt ggcggtgggc gcagcagtcg gtggggctgg cctgcggcac tgggctgaca
     1201 tgttggccaa ccccaggcgc cccattgccc agtggcactc tctgcggccc cctgacagag
     1261 ccaggccaat acctgcaccc tgagaattca tcgtgactga ctgacgatct gcctcgcgcg
     1321 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     1381 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     1441 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
     1501 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     1561 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1621 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1681 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1741 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1801 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1861 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1921 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     1981 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2041 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2101 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2161 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2221 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2281 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2341 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2401 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2461 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2521 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2581 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2641 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2701 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2761 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2821 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2881 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2941 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3001 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3061 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3481 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3541 tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc
     3601 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     3661 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     3721 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     3781 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     3841 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     3901 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     3961 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     4021 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     4081 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     4141 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     4201 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     4261 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     4321 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     4381 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     4441 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     4501 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     4561 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     4621 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4681 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4741 taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4801 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4861 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4921 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4981 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
     5041 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
     5101 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
     5161 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
     5221 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
     5281 ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     5341 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     5401 gcgcac
//
