LOCUS       sequence-260949-        4698 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pH measurement, nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4698)
  AUTHORS   Yaginuma H,  Okada Y
  TITLE     Live cell imaging of metabolic heterogeneity by quantitative 
            fluorescent ATP indicator protein, QUEEN-37C
  JOURNAL   bioRxiv 2021.10.08.463131
REFERENCE   2  (bases 1 to 4698)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2021.10.08.463131"
COMMENT     Sequence Label: pN1-NLS-pHlu
FEATURES             Location/Qualifiers
     source          1..4698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             546..566
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      579..588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1426..1547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1463..1482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1517..1536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1554..2009)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1691..1712)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1903..1922
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2036..2140
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2142..2499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2142..2333
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2165..2185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2350..2485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2534..3328
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2534..3328
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2588..2607)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3198..3217
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3516..3535)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3560..3607
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3936..4524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4425..4444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggcgccgaa aaaaaaacgc aaagtggatc caccggtcgc caccatggtg agcaagggtg
      601 aagagctgtt taccggcgtg gtgccgattc tggtggaact ggatggtgac gtgaacggcc
      661 acaaattcag cgtgtccggc gaaggcgagg gcgatgccac ctatggtaaa ctgaccctga
      721 agtttatctg caccaccggc aagctgccag tgccatggcc aaccctggtg accaccttta
      781 gctatggcgt gcagtgtttc tcccgctacc ccgatcacat gaaacgtcat gactttttca
      841 agagcgccat gccagaaggt tatgtgcagg agcgcaccat tttctttaaa gatgacggca
      901 attacaagac ccgtgcggaa gtgaaatttg agggcgatac cctggtgaac cgcattgaac
      961 tgaaaggcat cgacttcaag gatgacggta atattctggg ccacaagctg gaatataact
     1021 acaatgagca tctggtgtat atcatggcag ataaacagaa gaacggcacc aaagctattt
     1081 tccaggtgca ccataatatc gaggatggcg gtgtgcagct ggcagaccat tatcagcaga
     1141 acacccctat tggcgatggc cctgtgctgc tgccggacaa tcactacctg catacccagt
     1201 ctgctctgag taaagatccg aacgaaaagc gcgaccacat ggtgctgctg gagtttgtga
     1261 ccgccgcggg tatcacccat ggcatggacg agctgtacaa atgagcggcc gcgactctag
     1321 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1381 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1441 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1501 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     1561 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1621 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1681 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1741 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1801 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1861 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1921 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     1981 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2041 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2101 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2161 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2221 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2281 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2341 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2401 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2461 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2521 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2581 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2641 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2701 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2761 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2821 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2881 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2941 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3001 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3061 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3121 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3181 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3241 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3301 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3361 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3421 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3481 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3541 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     3601 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3661 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3721 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     3781 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3841 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3901 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3961 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4021 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4081 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4141 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4201 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4261 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4321 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4381 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4441 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4501 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4561 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4621 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     4681 cccatatatg gagttccg
//
