LOCUS       sequence-199533-        7724 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 7724)
  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 7724)
  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..7724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        75..454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        455..658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     609..629
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     703..722
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        703..721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      804..813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1445..1464
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3154..3171)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3160..3384
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3430..3858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3517..3536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3727..3748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3867..3887)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3872..4202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4053..4188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4115..4134
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4269..5063
                     /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(4323..4342)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4933..4952
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5237..5358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5274..5293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5328..5347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5407..5423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5407..5423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5420..5442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5431..5447
                     /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(5455..5461,5462..5479,5480..5485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5500..5521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5638..5655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5809..6397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5889..5908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6568..7359,7360..7428))
                     /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     7191..7210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7429..7533)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7608..7627)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
       61 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      121 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      181 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      241 aacgccaata gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca
      301 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      361 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      421 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      481 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      541 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      601 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      661 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
      721 ggagacccaa gctggctagc gtttaaactt aagcttggta ccgagctcgg atccactagt
      781 ccagtgtggt ggaattcacg cgtgccacca tggaacccac cgcgccaacc ggtcaggccc
      841 gggcggcggc caccaaactg tcggaagcgg tgggagccgc gctacaagag ccccagaggc
      901 agcggcgcct ggtgctggtc atcgtgtgcg ttgcactgtt actggacaac atgttgtaca
      961 tggtcatcgt gcccattgtt cccgactata tcgcccacat gcgcgggggc agcgagggcc
     1021 cgaccctggt cacgcgtagt aaaggagaag agaataacat ggccatcatc aaggagttca
     1081 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     1141 agggcgaggg ccgcccctac gaggcctttc agaccgctaa gctgaaggtg accaagggtg
     1201 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggtct
     1261 acattaagca cccagccgac atccccgact acttcaagct gtccttcccc gagggcttca
     1321 ggtgggagcg cgtgatgaac ttcgaggacg gcggcattat tcacgttaac caggactcct
     1381 ccctgcagga cggcgtgttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1441 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccgaggag cggatgtacc
     1501 ccgaggacgg cgccctgtac agcgagatca agaagaggct gaagctgaag gacggcggcc
     1561 actacgccgc cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1621 cctacatcgt cgacatcaag ttggacatcg tgtcccacaa cgaggactac accatcgtgg
     1681 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggatgaa ctatacaaac
     1741 cgaccctggt ctctgaggtg tgggaaccca ctctgccgcc gcccactctg gctaatgcca
     1801 gtgcctactt ggccaacacg tcggcgtccc cgacggctgc cgggtcggct cggtcaatcc
     1861 tgcgacctcg ctaccccaca gaaagcgaag atgtgaagat aggtgtgctg tttgcctcca
     1921 aggctatcct gcagcttctg gtgaacccct taagcgggcc tttcattgat cgcatgagct
     1981 acgacgtgcc gctgcttata ggcctgggcg tcatgttcgc ctccacagtc atgtttgcct
     2041 ttgcagaaga ctatgccacg ctcttcgctg cgcgcagtct acaaggcctg ggctcggcct
     2101 tcgcggacac gtctggcatt gccatgatcg ccgacaagta tcccgaggag cctgagcgca
     2161 gtcgtgccct gggcgtggcg ctagccttta ttagctttgg aagcctagtg gcgccaccgt
     2221 ttgggggcat cctctacgag ttcgcgggca agcgtgtacc ctttctagtg ctcgccgctg
     2281 tgtccctttt cgacgcgctc ctgctcctgg cggtggctaa gcccttctcg gctgcggctc
     2341 gggcgcgagc caacctgccg gtgggcacac ctatccatcg cctcatgcta gacccttaca
     2401 tcgctgtggt agccggcgcg ctcaccactt gtaacattcc ccttgcgttc ctcgagccca
     2461 ccatagccac gtggatgaag cacacaatgg ccgcatccga gtgggagatg ggcatggttt
     2521 ggctgccggc tttcgtgcca cacgtgttag gcgtctacct caccgtgcgc ctggcggcgc
     2581 gttatccaca cctgcagtgg ctgtacggcg ctctcgggct agcggtaatt ggagtgagct
     2641 cttgcgtcgt acctgcctgt cgctcattcg cgccgttagt ggtctcgctc tgcggactct
     2701 gcttcggcat cgcgttagtg gacacagcgc tcctacccac gctcgccttt ctggtggacg
     2761 tgcgccacgt atccgtctat ggcagtgtct atgccatagc tgacatctcc tattctgtgg
     2821 cctacgcgct cgggcccata gtggcaggcc acatcgttca ctctcttggc tttgagcagc
     2881 tcagcctggg catgggcctg gccaacctgc tctacgcacc agtccttctt cttttgcgca
     2941 atgtaggcct ccttacacgc tcgcgttcgg agcgcgatgt gttgcttgat gaaccgccgc
     3001 agggtctgta cgacgcggtg cgcctgcgtg aggtgcaggg caaggatggc ggcgaacctt
     3061 gtagcccacc tggccctttt gacgggtgcg aggacgacta caactattac tcccgcagct
     3121 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3181 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3241 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3301 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3361 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3421 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3481 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3541 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3601 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3661 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3721 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3781 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     3841 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3901 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3961 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4021 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4081 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4141 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4201 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4261 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4321 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4381 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4441 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4501 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4561 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4621 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4681 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4741 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4801 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4861 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4921 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4981 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5041 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5101 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5161 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5221 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5281 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5341 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5401 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5461 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5521 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5581 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5641 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5701 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5761 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5821 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5881 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5941 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6001 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6061 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6121 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6181 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6241 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6301 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6361 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6421 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6481 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6541 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6661 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6721 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7081 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7441 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7501 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7561 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7621 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7681 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccga
//
