LOCUS       sequence-391947-        6370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Synaptotagmin 5 with pH-sensitive 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6370)
  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 6370)
  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 Syt5
FEATURES             Location/Qualifiers
     source          1..6370
                     /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    3484..3605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3495..3514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3549..3568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3786..4241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3873..3892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4083..4104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4308..4327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4427..4449
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4487..4505)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4573..4677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4678..4746,4747..5538)
                     /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(4896..4915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5709..6297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6198..6217
                     /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 cggcggaccc ggggccaggg acgcgctctg tcaccaggcg ctgcagctgc tggccgagct
     2041 ctgtgcccga ggggccctgg agcacgacag ctgccaagat ttcatctacc acctgcggga
     2101 ccgtgccaga ccccggctcc gcgacccaga tatctctgtg agcctgctaa ccctcgtggt
     2161 tactgcctgt ggtctggccc tttttggtgt ctctcttttc gtgtcttgga aactgtgctg
     2221 ggttccgtgg cgagagcgag gcctgttctc tggtagcaaa gacaacaacc aagagcctct
     2281 taattacacc gacacagaga caaacgagca ggagaacagc gaggacttcc tagacccacc
     2341 tacaccctgc cctgactcct ccatgaagat aagccacact tcccccgaca ttcccctctc
     2401 cacccagcca gggggccagg agaactgtgc ccatgccgtc cgtgtgcagc gacaagtcac
     2461 agagccaaca ccatcagctc ggcataactc aatccgaaga cagctcaacc tgtcgaaccc
     2521 ggactttaat atccagcagc ttcagaggca ggagcagctg actgggattg gtagaattaa
     2581 accagagtta tacaaacaga ggtcactgga caacgacgat gggcggagga gtaacagcaa
     2641 agcctgcggg aaactgaact tcattttaaa atatgactgc gacttggaac agctcatcgt
     2701 gaagatccac aaagccgtca atctgcctgc caaggacttc tctgggactt cagatcctta
     2761 tgtcaagatc tacttgcttc ctgaccggaa aacaaaacac cagactaaag ttcacaggaa
     2821 gaccctgaac cctgtatttg atgaggtgtt tttatttcct gttcactaca atgaccttga
     2881 agctcggaag cttcacttct ctgtgtatga ctttgacagg ttctctcgcc atgacctgat
     2941 tggtcaggtg gtggtggacc acttcttcga cttggccgac ttccccaggg agtgcatcct
     3001 ttggaaggat atcgagtatg tcaccaacga taatgtggat ctgggcgagc ttatgttttc
     3061 actctgctat cttccaacgg ctggcaggct gaccatcacc atcataaaag caaggaattt
     3121 aaaggcgatg gacataacag gagcatcaga tccctacgtg aaagtctcac tgatgtgtga
     3181 cggccggaga ctgaagaaga ggaaaacctc caccaagagg aacacgctta atcccgtcta
     3241 caatgaagcc atagtcttcg acgtccctcc cgagagcatc gaccagatcc acttgtccat
     3301 agccgtcatg gactatgacc gtgtaggtca caatgaggtc attggcgtgt gccaagtagg
     3361 aaacgaggct gagcggctgg gcagagacca ctggagtgag atgctgtcat atcctcggaa
     3421 gcccatcgca cactggcact ctctgatgga gaaacgatga gcggccgctt cgagcagaca
     3481 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     3541 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3601 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     3661 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgggctggc
     3721 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3781 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3841 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3901 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3961 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4021 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4081 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4141 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4201 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     4261 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4321 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4381 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4441 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4501 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5941 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6001 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6061 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
     6361 cgacagatct
//
