LOCUS       sequence-383645-        5928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid encoding GST-tagged SYT1 C2AB domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5928)
  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 5928)
  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 C2AB
FEATURES             Location/Qualifiers
     source          1..5928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(46..51,52..67,68..74)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    82..98
                     /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             121..774
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     732..754
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             781..798
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1841..1863)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2008..2026)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2094..2198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2199..2267,2268..3059)
                     /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(2417..2436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3230..3818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3719..3738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3972..3989
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4062..4139
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4140..5222
                     /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(4159..4178)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5271..5276,5277..5294,5295..5301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5309..5325
                     /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     5333..5349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5333..5349
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5345..5872
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5365..5382)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5365..5381)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5374..5396)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac aattaatcat
       61 cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagtattc
      121 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt
      181 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa
      241 tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat
      301 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac
      361 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg
      421 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt
      481 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa
      541 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat
      601 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa
      661 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca
      721 tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat
      781 ctggttccgc gtggatccat gggaggaaag aacgccatta acatgaaaga cgtgaaagac
      841 ttagggaaga ccatgaagga tcaggccctt aaggatgacg atgctgaaac cggactgact
      901 gatggagaag aaaaggaaga gcccaaggaa gaggagaaac tgggaaagct ccaatattca
      961 ctggactatg acttccagaa taaccagctg ttggtgggaa tcatccaggc tgctgaactg
     1021 cccgccctgg acatgggggg tacatccgat ccatacgtca aagtcttcct gctgcctgac
     1081 aaaaagaaga aatttgagac taaagtccac cggaaaaccc tcaatccagt cttcaatgaa
     1141 caatttactt tcaaggtacc ctactcggaa ttaggtggca aaaccctggt gatggctgtg
     1201 tatgactttg atcgcttctc caagcacgac atcatcggag agttcaaagt tcctatgaac
     1261 accgtggatt ttggccatgt gaccgaggag tggcgcgatc tccagagcgc tgagaaagaa
     1321 gagcaagaga aactgggtga catctgcttc tccctccgct acgtccctac tgccggcaaa
     1381 ctgactgttg tcattctgga agccaagaac ctgaagaaga tggatgtggg tggcttatct
     1441 gatccctacg tgaagattca cctgatgcag aacggtaaga ggctgaagaa gaaaaagacg
     1501 acgattaaga agaacacact caacccctac tacaacgagt ccttcagctt tgaagttccg
     1561 ttcgagcaaa tccagaaagt gcaagtggtg gtaactgttt tggactatga caagattggc
     1621 aagaacgacg ccatcggcaa agtcttcgtt ggttacaaca gcactggggc ggagctgcga
     1681 cactggtcag acatgctggc caacccccgg cgacccatcg cacagtggca cactctgcag
     1741 gtagaggagg aggttgatgc catgctggct gtcaagaagt aagaattcat cgtgactgac
     1801 tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1861 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1921 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     1981 gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     2041 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2101 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2161 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2221 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2281 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2341 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2401 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     2461 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2521 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2581 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2641 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2701 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     2761 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2821 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2881 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2941 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3001 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3061 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3121 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3181 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3241 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3301 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3361 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3421 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3481 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3541 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3601 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3661 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3721 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3781 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3841 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3901 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3961 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     4021 tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc gaatggcgca
     4081 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     4141 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     4201 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     4261 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     4321 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     4381 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     4441 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     4501 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     4561 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     4621 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     4681 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     4741 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     4801 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     4861 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     4921 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
     4981 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     5041 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     5101 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     5161 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5221 gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc tttacacttt
     5281 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5341 agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5401 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5461 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5521 gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct
     5581 tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc
     5641 catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa
     5701 tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca
     5761 gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt gcaccaatgc
     5821 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
     5881 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgtttttt
//
