LOCUS       sequence-207582-        7357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synaptic vesicle protein VGluT1 fused to the SNAPtag domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7357)
  AUTHORS   Martineau M, Somasundaram A, Grimm JB, Gruber TD, Choquet D, Taraska
            JW, Lavis LD, Perrais D
  TITLE     Semisynthetic fluorescent pH sensors for imaging exocytosis and 
            endocytosis.
  JOURNAL   Nat Commun. 2017 Nov 10;8(1):1412. doi: 10.1038/s41467-017-01752-5.
  PUBMED    29123102
REFERENCE   2  (bases 1 to 7357)
  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"; journalName: "Nat 
            Commun."; date: "2017-11-10"; volume: "8(1):1412. doi"; pages: " 
            10.1038/s41467-017-01752-5"
COMMENT     Sequence Label: VGluT1-SNAPtag
FEATURES             Location/Qualifiers
     source          1..7357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..657
                     /label=chicken beta-actin promoter
     intron          658..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1840..1849
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2200..2742
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for mammalian expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     CDS             complement(3809..3820)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4375..4394)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4440..4495
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4494..4513)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4856..4872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4856..4872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4869..4891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4880..4896
                     /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)."
     promoter        complement(join(4904..4910,4911..4928,4929..4934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4949..4970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5029..5224
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5075..5210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5230..5364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5267..5286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5321..5340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5432..5449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5603..6191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5683..5702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6362..7153,7154..7222))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6985..7004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7223..7327)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
      541 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      661 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      781 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      841 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
      901 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      961 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgg cacgagcgga
     1741 gctgcggggc cgggccgggc cggggcggac cccgggatcc cggacgcggc cgcccgggcc
     1801 cgcgggcggg gggattggca ggggacccgc gtgggcacag ccaccatgga gttccggcag
     1861 gaggagtttc ggaagctggc ggggcgcgcc ctggggaggc tgcaccggtt actggagaag
     1921 cggcaggaag gcgcggagac attggagctg agcgccgacg ggcgcccagt gaccacacac
     1981 acgcgggacc cgccggtggt ggactgcact tgctttggcc tccctcgccg ctacatcatc
     2041 gcgatcatga gcggtctggg tttctgcatc agctttggca tccgctgcaa cctgggcgtg
     2101 gccatcgtat ccatggtcaa caacagtaca acccaccgtg ggggccacgt ggtgtccggt
     2161 agcactagcg gcggaagcgg cgggacaggt ggatccggag acaaagactg cgaaatgaag
     2221 cgcaccaccc tggatagccc tctgggcaag ctggaactgt ctgggtgcga acagggcctg
     2281 cacgagatca agctgctggg caaaggaaca tctgccgccg acgccgtgga agtgcctgcc
     2341 ccagccgccg tgctgggcgg accagagcca ctgatgcagg ccaccgcctg gctcaacgcc
     2401 tactttcacc agcctgaggc catcgaggag ttccctgtgc cagccctgca ccacccagtg
     2461 ttccagcagg agagctttac ccgccaggtg ctgtggaaac tgctgaaagt ggtgaagttc
     2521 ggagaggtca tcagctacca gcagctggcc gccctggccg gcaatcccgc cgccaccgcc
     2581 gccgtgaaaa ccgccctgag cggaaatccc gtgcccattc tgatcccctg ccaccgggtg
     2641 gtgtctagct ctggcgccgt ggggggctac gagggcgggc tcgccgtgaa agagtggctg
     2701 ctggcccacg agggccacag actgggcaag cctgggctgg gtcctgcagg tataggcgcg
     2761 ccaggatccc tcgaggctag cagcggaggc acaggtggca gcggcggaac cggaggctcc
     2821 ggtggcaccg gagtgcagaa agcccagttc aactgggatc cagagactgt cggcctcata
     2881 catggctcct ttttctgggg gtacattgtc actcagattc ctggaggatt tatctgccaa
     2941 aaattcgcag ccaacagggt ctttggcttt gccattgtgg ctacctccac cctaaatatg
     3001 ttgatccctt cagcagcccg tgttcactat ggctgtgtca tcttcgtgag gatccttcag
     3061 ggattggtgg agggggtcac ataccctgct tgccatggca tctggagcaa atgggcccct
     3121 cccttagaac ggagtcggct ggcgacgaca gccttttgcg gttcctatgc cggggcagtg
     3181 gttgccatgc ctctggctgg ggtcctggta cagtattcag gatggagttc tgtcttctat
     3241 gtctatggca gcttcgggat cttttggtac ctgttctggt tgcttgtctc ctacgagtca
     3301 cctgcactac accccagcat ctccgaggag gagcgcaaat acattgagga tgccatcgga
     3361 gaaagcgcca agctcatgaa ccctgttacg aagtttaaca caccctggag gcgcttcttt
     3421 acctccatgc cggtctatgc catcattgtc gccaactttt gccgcagctg gactttctac
     3481 ctgctcctca tctcccagcc cgcctacttt gaagaagtgt tcggctttga gatcagcaag
     3541 gtgggactgg tgtcggcact gcctcacctt gtcatgacta tcatcgtacc catcggaggc
     3601 cagatcgccg acttcctgcg cagtcgtcat ataatgtcca cgaccaatgt gcgaaagctg
     3661 atgaactgcg ggggtttcgg gatggaagct acgctgctgc tggtggtcgg atactcacac
     3721 tccaagggcg tggccatctc cttcctggtc ctggctgtgg gcttcagtgg ctttgctatc
     3781 tctgggttta acgtgaacca cttggacatc gcccctcgat atgccagcat cttgatgggc
     3841 atttccaatg gcgtgggcac actgtctggg atggtgtgcc ccatcatcgt gggtgcaatg
     3901 accaagcaca agacgcggga ggagtggcag tacgtgttcc tcatagcctc cctggtgcac
     3961 tatggaggtg tcatcttcta tggggtcttt gcttcgggag agaaacagcc gtgggcagag
     4021 ccggaggaga tgagcgagga gaagtgtggc tttgttggcc acgaccagct ggctggcagt
     4081 gacgaaagtg aaatggaaga cgaggttgag cccccggggg caccccccgc acctccgcct
     4141 tcctacgggg ccacacacag cacagttcag cctccaaggc ccccaccccc tgtccgggac
     4201 tactgaccac gtgcctccca ctggtgggca gtttccagga cctccactcg atacacctct
     4261 agcctaaacg gcagtgtcga ggaaccccac tcctctcctg cctcaggtgc aggctgccta
     4321 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     4381 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     4441 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     4501 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     4561 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     4621 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     4681 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     4741 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     4801 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     4861 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4921 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4981 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     5041 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5101 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5161 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5221 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5281 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5341 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     5401 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5461 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5521 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5581 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5641 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5701 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5761 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5821 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5881 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5941 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6001 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6061 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6121 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6301 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6361 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6421 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6481 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6541 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6601 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6661 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6721 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6781 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6841 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6901 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6961 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7021 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7081 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7141 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7201 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7321 ttccgcgcac atttccccga aaagtgccac ctgggtc
//
