LOCUS       sequence-393103-        6250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Synaptotagmin 17 with pH-sensitive 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  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 6250)
  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 Syt17
FEATURES             Location/Qualifiers
     source          1..6250
                     /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"
     CDS             1058..1147
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             1166..1876
                     /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(1193..1221)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1816..1836
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3364..3485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3375..3394)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3429..3448
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3666..4121
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3753..3772)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3963..3984
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4188..4207)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4307..4329
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4367..4385)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4453..4557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4558..4626,4627..5418)
                     /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(4776..4795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5589..6177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6078..6097
                     /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 ggctagcatg gacagcaaag gttcgtcgca
     1081 gaaagggtcc cgcctgctcc tgctgctggt ggtgtcaaat ctactcttgt gccagggtgt
     1141 ggtctccgat atcggtacca cgcgtagtaa aggcgaagaa cttttcactg gagttgtccc
     1201 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
     1261 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
     1321 acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcaat gcttttcaag
     1381 atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
     1441 acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa
     1501 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
     1561 tggaaacatt cttggacaca aattggaata caactataac gatcaccagg tgtacatcat
     1621 ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca acattgaaga
     1681 tggaggcgtt caactagcag accattatca acaaaatact ccaattggcg atgggcccgt
     1741 ccttttacca gacaaccatt acctgtttac aacttctact ctttcgaaag atcccaacga
     1801 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
     1861 ggatgaacta tacaaagaat tctcaggcgg aagcggaggc tccggcggga gcggcggagc
     1921 gtacatccag ttggaaccac tgaacgaggg ctttctttcc agaatctctg atgtgctgct
     1981 gtgtggatgg acttgtcaac actgctgtca gaggtgctat gagtccagct gctgccagtc
     2041 tagtgaagat gaagtggaga tcttgggacc tttccctgcc cagactcctc catggctgat
     2101 ggccagccgg agcaatgaca aggatggtga ctctgtgcac acagccagcg atgttccttt
     2161 gaccccaagg accaactccc ctgatgggag acgctcatcc tcagacacat ccaagtccac
     2221 atacagcctg actcggagaa tctccagtct ggattcccgg cgccccagct ccccactcat
     2281 tgacatcaaa cctattgagt tcggtgttct cagtgccaag aaggagccca tccagccatc
     2341 tgtgctgcgc cgaacctaca cccctgatga ctacttcagg aagtttgagc cccgacttta
     2401 ctcccttgac tcgaatcttg atgacgtaga ctctctgacc gatgaggaga taatgtccaa
     2461 gtaccagctg ggcatgctgc acttcagcac ccagtatgac ctgctgcaca accacctcac
     2521 cgtgagggtg atcgaggccc gggacctgcc gcctccaatc tcccatgatg gctcccgcca
     2581 ggacatggcc cattccaacc cctacgtcaa gatctgcctc ctgcctgacc agaagaactc
     2641 caagcagacg ggagtcaaac gcaagaccca gaagcctgtg tttgaggaac gctacacctt
     2701 cgagatcccc ttcctagaag cccagaggag gaccctgctg ctcactgtgg tggattttga
     2761 caaattctcc cgccactgtg tgattgggaa agtagcggtg cctttgtgtg aggtggactt
     2821 ggtgaagggc ggccactggt ggaaagctct gatccccagc tctcagaatg aagtggagct
     2881 gggagagctg ctgctttctc taaactatct cccaagtgct ggcagactga atgtggacat
     2941 cattcgagcc aagcaactcc tgcagacaga tgtgagccag ggttcagacc catttgtgaa
     3001 aatccagctg gtacacggac tcaagcttgt gaaaactaag aagacatcat tcttacgagg
     3061 cacaatcgat cctttctaca atgaatcctt cagcttcaaa gttccccagg aagaactaga
     3121 aaacgccagc ctagtattca cagtgttcgg tcacaacatg aaaagcagca acgacttcat
     3181 cgggaggatc gtcatcggcc agtattcctc cggcccctcg gaatccaacc actggcggcg
     3241 aatgctcaac acacaccgca ctgctgtgga acagtggcat agcctgaggt cccgggctga
     3301 gtgtgaccga gtgtctccgg cctcgctgga ggtgacctga gcggccgctt cgagcagaca
     3361 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     3421 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3481 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     3541 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgggctggc
     3601 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3661 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3721 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3781 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3841 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3901 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3961 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4021 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4081 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     4141 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4201 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4261 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4321 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4381 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4441 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4501 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4561 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4621 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4681 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4741 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4801 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4861 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4921 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4981 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5041 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5101 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5161 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5221 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5281 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5341 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5401 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5461 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5701 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
     6241 cgacagatct
//
