LOCUS       sequence-142703-        3432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ratiometric pHluorin2 with a gpi linkage sequence in a middle entry 
            clone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3432)
  AUTHORS   Stawicki TM, Owens KN, Linbo T, Reinhart KE, Rubel EW, Raible DW
  TITLE     The zebrafish merovingian mutant reveals a role for pH regulation in
            hair cell toxicity and function.
  JOURNAL   Dis Model Mech. 2014 Jul;7(7):847-56. doi: 10.1242/dmm.016576.
  PUBMED    24973752
REFERENCE   2  (bases 1 to 3432)
  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: "10.1242/dmm"; 
            journalName: "Dis Model Mech"; date: "2014-07"; volume: "7"; issue: 
            "7"; pages: "847-56"
COMMENT     Sequence Label: pME phluorin2-gpi
FEATURES             Location/Qualifiers
     source          1..3432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(797..818)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1058..1077)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1405..1426
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1557..1656)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1673..1692)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1674..1692)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1697..1713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1697..1713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1815..1834)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1826..2635
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1893..1912)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2782..3370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3271..3290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctaa atggctcagc ggatgacaac acagctgctg ctccttctag tgtgggtggc
      721 tgtagtaggg gaggctcaga caaggattgc aatggtgagc aagggcgagg agctgttcac
      781 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      841 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      901 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgagct acggcgtgca
      961 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1021 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1081 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1141 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acgagcacct
     1201 ggtgtacatc atggccgaca agcagaagaa cggcaccaag gccatcttcc aggtgcacca
     1261 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1321 cgacggcccc gtgctgctgc ccgacaacca ctacctgcac acccagtccg ccctgagcaa
     1381 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1441 cactcacggc atggacgagc tgtacaaggc tgcagccatg agtggggctg ggccctgggc
     1501 agcctggcct ttcctgctta gcctggccct aatgctgctg tggctgctca gctgataccc
     1561 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     1621 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     1681 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     1741 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     1801 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     1861 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1921 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     1981 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2041 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2101 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2161 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2221 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2281 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2341 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2401 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2461 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2521 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2581 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2641 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     2701 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     2761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3421 tgctcacatg tt
//
