LOCUS       sequence-391951-        6106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Synaptotagmin 7 with pH-sensitive 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6106)
  AUTHORS   Dean C, Dunning FM, Liu H, Bomba-Warczak E, Martens H, Bharat V, 
            Ahmed S, Chapman ER
  TITLE     Axonal and dendritic synaptotagmin isoforms revealed by a 
            pHluorin-syt functional screen.
  JOURNAL   Mol Biol Cell. 2012 May;23(9):1715-27. doi: 10.1091/mbc.E11-08-0707.
            Epub 2012 Mar 7.
  PUBMED    22398727
REFERENCE   2  (bases 1 to 6106)
  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: 
            "10.1091/mbc.E11-08-0707"; journalName: "Mol Biol Cell"; date: 
            "2012-05"; volume: "23"; issue: "9"; pages: "1715-27"
COMMENT     Sequence Label: pCI pHluorin Syt7
FEATURES             Location/Qualifiers
     source          1..6106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     718..742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           1076..1118
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1127..1216
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             1235..1945
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1262..1290)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1885..1905
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3220..3341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3231..3250)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3285..3304
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3522..3977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3609..3628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3819..3840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4044..4063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4163..4185
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4223..4241)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4309..4413
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4414..4482,4483..5274)
                     /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     complement(4632..4651)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5445..6033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5934..5953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata ggctagcctc gagaattcga tatcgcttgt
     1081 tctttttgca gaagctcaga ataaacgctc aactttggca gataccatgg acagcaaagg
     1141 ttcgtcgcag aaagggtccc gcctgctcct gctgctggtg gtgtcaaatc tactcttgtg
     1201 ccagggtgtg gtctccgata tcggtaccac gcgtagtaaa ggcgaagaac ttttcactgg
     1261 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag
     1321 tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac
     1381 tggaaaacta cctgttcctt ggccaacact tgtcactact ttaacttatg gtgttcaatg
     1441 cttttcaaga tacccagatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga
     1501 aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtgc
     1561 tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt
     1621 taaagaagat ggaaacattc ttggacacaa attggaatac aactataacg atcaccaggt
     1681 gtacatcatg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttagacacaa
     1741 cattgaagat ggaggcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     1801 tgggcccgtc cttttaccag acaaccatta cctgtttaca acttctactc tttcgaaaga
     1861 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1921 acatggcatg gatgaactat acaaagaatt ctcaggcgga agcggaggct ccggcgggag
     1981 cggcggatac cgggacccgg aggcggccag cccaggggca cctacccgcg atgtcctgct
     2041 ggtctctgca atcatcaccg tcagccttag cgtcactatc gtcctctgcg gcctgtgcca
     2101 ctggtgtcag cgcaaactgg gcaaacgcta caagaattcc ttggagacgg tgggcacgcc
     2161 agactcgggg cgtgggcgcg gtgagaagaa agccatcaag ttgcctgcag gagggaaggc
     2221 tgtgaataca gccccagtgc ccggccagac gccacacgat gagtctgacc gcagaacgga
     2281 gacccgttcc tctgtctcgg acctcgtcaa ctcccttacc agcgagatgc tcatgctctc
     2341 cccaggttct gaggaggatg aggcccacga gggctgcagc cgagagaacc tgggccgaat
     2401 ccagttcagt gttggctaca acttccaaga gtccacactc accgtgaagg tcatgaaggc
     2461 ccaagagctg ccagccaagg acttcagcgg tactagtgac ccctttgtca agatctacct
     2521 gctacccgac aagaagcaca aactggagac caaggtgaag cggaagaatc taaacccgca
     2581 ctggaatgag acctttctat ttgaagggtt tccctacgag aaagtggtgc agagggtcct
     2641 ctacctccag gtcctggatt atgaccgttt cagccgcaat gaccccattg gggaggtgtc
     2701 catccctctg aacaaggtgg acctgaccca gatgcagacc ttctggaagg atctgaagcc
     2761 atgcagcgat gggagtggga gccgagggga gctgctcttg tccctctgct acaacccctc
     2821 tgccaactcc atcatcgtga acatcatcaa agctcgaaac ctcaaagcca tggacatcgg
     2881 gggcacatca gacccctatg tgaaggtgtg gctgatgtat aaagacaagc gggtagagaa
     2941 aaagaagact gtgacaaaga agaggaacct gaaccccatc ttcaatgagt ctttcgcctt
     3001 cgacataccc acggagaagc tgagggagac cacgatcatc atcactgtca tggacaaaga
     3061 caagctcagc cgcaacgacg tcatcggcaa gatctacctg tcctggaaga gcggaccagg
     3121 tgaagtgaaa cactggaagg acatgatcgc tcgtccccgg cagcctgtgg cccagtggca
     3181 ccagctgaaa gcctgagcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     3241 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3301 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3361 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     3421 acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa tagcgaagag gcccgcaccg
     3481 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     3541 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3601 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3661 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3721 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3781 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3841 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3901 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3961 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4021 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4081 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4141 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4201 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4261 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4321 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4381 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4441 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4501 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4561 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4621 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4681 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4741 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4801 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4861 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4921 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4981 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5041 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5101 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5161 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5221 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5281 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5341 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5401 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5461 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5521 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5581 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5641 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5701 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5761 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5821 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5881 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5941 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6001 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6061 ttcctggcct tttgctggcc ttttgctcac atggctcgac agatct
//
