LOCUS       sequence-414386-        6423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Synaptotagmin 6 with pH-sensitive 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  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 6423)
  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 Syt6
FEATURES             Location/Qualifiers
     source          1..6423
                     /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           1069..1111
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1120..1209
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             1228..1938
                     /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(1255..1283)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1878..1898
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3537..3658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3548..3567)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3602..3621
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3839..4294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3926..3945)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4136..4157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4361..4380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4480..4502
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4540..4558)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4626..4730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4731..4799,4800..5591)
                     /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(4949..4968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5762..6350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6251..6270
                     /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 gagaattcct tgttcttttt
     1081 gcagaagctc agaataaacg ctcaactttg gcagatacca tggacagcaa aggttcgtcg
     1141 cagaaagggt cccgcctgct cctgctgctg gtggtgtcaa atctactctt gtgccagggt
     1201 gtggtctccg atatcggtac cacgcgtagt aaaggcgaag aacttttcac tggagttgtc
     1261 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     1321 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     1381 ctacctgttc cttggccaac acttgtcact actttaactt atggtgttca atgcttttca
     1441 agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     1501 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc
     1561 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
     1621 gatggaaaca ttcttggaca caaattggaa tacaactata acgatcacca ggtgtacatc
     1681 atggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattagaca caacattgaa
     1741 gatggaggcg ttcaactagc agaccattat caacaaaata ctccaattgg cgatgggccc
     1801 gtccttttac cagacaacca ttacctgttt acaacttcta ctctttcgaa agatcccaac
     1861 gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
     1921 atggatgaac tatacaaaga attctcaggc ggaagcggag gctccggcgg gagcggcgga
     1981 agcggagttt ggggggccgg agggcctcgg tgccaggcgg cgctcgcggt cctggcttct
     2041 ctgtgccggg cccggccgcc ccctctcggg ctggacgttg agacttgtcg gagcttcgag
     2101 ctgcagtccc cggagcagag tccaagcgcg gcagactcag gcacctctgt cagcctcctc
     2161 gccgtggtag ttattgtgtg tggcgtggcc ctggtggcag tttttctctt tctcttttgg
     2221 aagctgtgct ggatgccctg gaggaagaaa gaggcctcca gcccttcttc tgctaaccct
     2281 gcttcagaga ctctccagag ccccagctcc agaggcaaca tggcggataa gctgaaggac
     2341 cccagtgccc tgggcttcct ggaggcagct gtgaagatca gccacacttc cccagacatc
     2401 ccagccgagg tgcagatgtc ggtcaaagag cacatcatgc gccacacaaa gctgcagcga
     2461 cagaccacag agccagcatc ctccaccagg cacacatcct tcaagcgcca cctgccacga
     2521 cagatgcatg tctccagcgt ggactatggc aatgagctgc cgccggctgc cgccgagcag
     2581 cccaccagta ttggccgtat caagcctgag ctttacaagc agaagtcagt ggatggggat
     2641 gatgccaagt cggaggccgc caagagctgt gggaagatca acttcagcct ccgctatgac
     2701 tatgaaagcg agacgctgat tgtgcgcatc ctgaaggcct ttgacctccc tgccaaggac
     2761 ttttgcggaa gttctgaccc ttatgttaag atctacctcc tgcctgaccg caagtgcaag
     2821 ctgcagaccc gggtgcaccg caagaccctg aaccccacct ttgatgagaa cttccacttc
     2881 cccgtgccct acgaggagct ggctgaccgc aagctgcatc tcagtgtctt tgactttgac
     2941 cgcttctccc gccatgacat gattggggag gtcatcctgg acaatctctt tgaggcctct
     3001 gacctgtccc gggagacctc catctggaag gacatccagt acgctactag tgaaagtgtg
     3061 gacttgggag agatcatgtt ctccctctgc tacctgccca cagcaggcag gctcaccctc
     3121 acagtgatca agtgtcgaaa tctcaaagca atggacatca caggctactc agatccctac
     3181 gtcaaagtat ccctactgtg tgatggacgg aggctgaaaa agaagaaaac gaccataaaa
     3241 aagaacaccc taaaccccgt ctacaacgag gccatcatct ttgacatccc cccagaaaac
     3301 atggaccaag tgagcctgct catctccgtt atggactatg atagagttgg ccacaacgag
     3361 atcataggag tctgtagagt ggggatcaat gctgaaggcc ttggcaggga ccactggaat
     3421 gagatgttgg cgtacccacg aaagcccatt gcgcactggc actccttggt ggaggtaaag
     3481 aaatccttca aagaggggac ccctcggttg tgagcggccg cttcgagcag acatgataag
     3541 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3601 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3661 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     3721 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgggct ggcgtaatag
     3781 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     3841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3901 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3961 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4021 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4141 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4201 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4261 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     4321 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     4381 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4441 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4501 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4561 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4621 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4681 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4741 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4801 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4861 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4921 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4981 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5041 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5101 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5161 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5221 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5281 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5341 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5401 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5461 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5521 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5581 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5641 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5701 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5761 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5821 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5881 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5941 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6001 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6061 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6121 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6181 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6241 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6301 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6361 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga
     6421 tct
//
