LOCUS       sequence-391950-        6466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Synaptotagmin 10 with pH-sensitive 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6466)
  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 6466)
  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 Syt10
FEATURES             Location/Qualifiers
     source          1..6466
                     /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"
     RBS             2577..2588
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     polyA_signal    3580..3701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3591..3610)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3645..3664
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3882..4337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3969..3988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4179..4200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4404..4423)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4523..4545
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4583..4601)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4669..4773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4774..4842,4843..5634)
                     /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(4992..5011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5805..6393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6294..6313
                     /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 cggcggaagt ttccgcaagg aggacggtgt gagcagcctg tgccagaagg cgctgcacat
     2041 catcaccgag ctgtgcttcg cgggtcaggt ggagtgggac aagtgttcgg gcatctttcc
     2101 agccgatagg agcggccagg ggggaggtgg cacagatatt tcagtcagcc tgttagcggt
     2161 tgttgtaagc ttctgtggcc tggccttgct ggttgtctcc ctttttgtct tctggaagtt
     2221 gtgctggcca tgctggaaga gcaagcttgt ggcccccaac ctcagtgtcc ttccacaaag
     2281 catctcaagt gctccaactg aagtttttga gactgaagag aaaaaagaag tggaagaaaa
     2341 tgaaaagcca gcacccaaag ccattgagcc tgcaataaaa atcagccaca catcccccga
     2401 catcccagca gaagtgcaga cagctttaaa ggagcacttg attaaacatg ctcgagtgca
     2461 aaggcaaaca accgagccta cgtcttcatc ccgccacaac tccttcagga gacacctacc
     2521 aaggcaaatg aatgtctcca gcgtcgactt tagcgtgggc actgaaccca tcttacaaag
     2581 aggagaaacg agaacgagca tcgggaggat aaaaccagaa ctctacaagc agaaatcggt
     2641 tgactccgag ggcaacagaa aagacgacgt caagacctgt gggaaactta actttgcgct
     2701 ccagtatgat tatgaaaatg aactcctagt tgttaaaatt atcaaagctt tagatctccc
     2761 agctaaagat ttcacaggaa cttctgatcc ttatgtgaag atctatcttc ttccggatag
     2821 gaagaagaaa tttcagaccc gcgtgcaccg gaagacttta aaccctctgt ttgatgagct
     2881 atttcagttt ccggtggttt acgatcagct aagcaaccgc aaactacact ttagtatata
     2941 tgattttgac agattttcca gacacgacat gattggggaa gtgattcttg ataatttatt
     3001 tgaagtctct gatctctcca gggaagccac agtatggaaa gatatccact gtgctaccac
     3061 agaaagcata gacctggggg aaatcatgtt ctccctttgc tatttgccaa ctgctggccg
     3121 tatgaccttg acagtgatca agtgcagaaa tctgaaggca atggacatca ctgggtcatc
     3181 agatccttat gtcaaagtgt ctctgatgtg tgagggccga agacttaaaa aacggaaaac
     3241 aactacaaag aaaaatactc tcaatcccgt gtacaacgaa gccattatct tcgatatccc
     3301 tccagagaat gtggaccagg tcagcctgtg cattgcagtc atggattacg acagggtggg
     3361 gcacaatgag gtcataggcg tgtgtcgaac tggactagac gctgagggtc ttgggagaga
     3421 ccattggaac gagatgctgg cctatcaccg aaagccaatc acacactggc accccttgct
     3481 ggagttacct ggccgggcca ccagctttga tagtcaaggg tcctgttctt ctcccaggcc
     3541 accgtccaca ccataagcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     3601 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3661 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3721 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     3781 acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa tagcgaagag gcccgcaccg
     3841 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4081 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4201 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4321 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4381 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4441 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4501 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4561 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4621 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4741 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4801 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4861 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4921 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4981 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5041 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5101 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5161 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5221 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5281 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5341 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5401 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5461 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5521 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5581 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5641 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5701 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5761 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5821 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5881 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5941 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6001 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6061 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6121 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6181 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6241 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6301 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6361 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6421 ttcctggcct tttgctggcc ttttgctcac atggctcgac agatct
//
