LOCUS       sequence-215606-        7723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VAChT tagged with pH-sensitive red fluorescent protein pHuji.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7723)
  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 7723)
  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: VAChT-pHuji
FEATURES             Location/Qualifiers
     source          1..7723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      730..739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1371..1390
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3080..3097)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3086..3310
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3356..3784
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3443..3462)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3653..3674
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3793..3813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3798..4127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3978..4113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4040..4059
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4194..4988
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4248..4267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4858..4877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5162..5283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5199..5218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5253..5272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5332..5348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5332..5348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5345..5367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5356..5372
                     /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(5380..5386,5387..5404,5405..5410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5425..5446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5563..5580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5734..6322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5814..5833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6493..7284,7285..7353))
                     /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     7116..7135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7354..7458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7533..7552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttcacgcgtg ccaccatgga acccaccgcg ccaaccggtc aggcccgggc ggcggccacc
      781 aaactgtcgg aagcggtggg agccgcgcta caagagcccc agaggcagcg gcgcctggtg
      841 ctggtcatcg tgtgcgttgc actgttactg gacaacatgt tgtacatggt catcgtgccc
      901 attgttcccg actatatcgc ccacatgcgc gggggcagcg agggcccgac cctggtcacg
      961 cgtagtaaag gagaagagaa taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1021 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1081 ccctacgagg cctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc
     1141 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggtctacat taagcaccca
     1201 gccgacatcc ccgactactt caagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg
     1261 atgaacttcg aggacggcgg cattattcac gttaaccagg actcctccct gcaggacggc
     1321 gtgttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1381 cagaagaaga ccatgggctg ggaggcctcc gaggagcgga tgtaccccga ggacggcgcc
     1441 ctgtacagcg agatcaagaa gaggctgaag ctgaaggacg gcggccacta cgccgccgag
     1501 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta catcgtcgac
     1561 atcaagttgg acatcgtgtc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     1621 gccgagggcc gccactccac cggcggcatg gatgaactat acaaaccgac cctggtctct
     1681 gaggtgtggg aacccactct gccgccgccc actctggcta atgccagtgc ctacttggcc
     1741 aacacgtcgg cgtccccgac ggctgccggg tcggctcggt caatcctgcg acctcgctac
     1801 cccacagaaa gcgaagatgt gaagataggt gtgctgtttg cctccaaggc tatcctgcag
     1861 cttctggtga accccttaag cgggcctttc attgatcgca tgagctacga cgtgccgctg
     1921 cttataggcc tgggcgtcat gttcgcctcc acagtcatgt ttgcctttgc agaagactat
     1981 gccacgctct tcgctgcgcg cagtctacaa ggcctgggct cggccttcgc ggacacgtct
     2041 ggcattgcca tgatcgccga caagtatccc gaggagcctg agcgcagtcg tgccctgggc
     2101 gtggcgctag cctttattag ctttggaagc ctagtggcgc caccgtttgg gggcatcctc
     2161 tacgagttcg cgggcaagcg tgtacccttt ctagtgctcg ccgctgtgtc ccttttcgac
     2221 gcgctcctgc tcctggcggt ggctaagccc ttctcggctg cggctcgggc gcgagccaac
     2281 ctgccggtgg gcacacctat ccatcgcctc atgctagacc cttacatcgc tgtggtagcc
     2341 ggcgcgctca ccacttgtaa cattcccctt gcgttcctcg agcccaccat agccacgtgg
     2401 atgaagcaca caatggccgc atccgagtgg gagatgggca tggtttggct gccggctttc
     2461 gtgccacacg tgttaggcgt ctacctcacc gtgcgcctgg cggcgcgtta tccacacctg
     2521 cagtggctgt acggcgctct cgggctagcg gtaattggag tgagctcttg cgtcgtacct
     2581 gcctgtcgct cattcgcgcc gttagtggtc tcgctctgcg gactctgctt cggcatcgcg
     2641 ttagtggaca cagcgctcct acccacgctc gcctttctgg tggacgtgcg ccacgtatcc
     2701 gtctatggca gtgtctatgc catagctgac atctcctatt ctgtggccta cgcgctcggg
     2761 cccatagtgg caggccacat cgttcactct cttggctttg agcagctcag cctgggcatg
     2821 ggcctggcca acctgctcta cgcaccagtc cttcttcttt tgcgcaatgt aggcctcctt
     2881 acacgctcgc gttcggagcg cgatgtgttg cttgatgaac cgccgcaggg tctgtacgac
     2941 gcggtgcgcc tgcgtgaggt gcagggcaag gatggcggcg aaccttgtag cccacctggc
     3001 ccttttgacg ggtgcgagga cgactacaac tattactccc gcagctagtc tagagggccc
     3061 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3121 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3181 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3241 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3301 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3361 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3421 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3481 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3541 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3601 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3661 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3721 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3781 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3841 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3901 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3961 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4021 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4081 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4141 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4201 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4261 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4321 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4381 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4441 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4501 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4561 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4621 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4681 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4741 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4801 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4861 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4921 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4981 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5041 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5101 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5161 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5221 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5281 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5341 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5401 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5461 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5521 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5581 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5641 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5701 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5761 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5821 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5881 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5941 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6001 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6061 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6601 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6781 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7501 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7561 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7621 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     7681 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
