LOCUS       sequence-218932-        6552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blue ATP sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6552)
  AUTHORS   Arai S, Kriszt R, Harada K, Looi LS, Matsuda S, Wongso D, Suo S, 
            Ishiura S, Tseng YH, Raghunath M, Ito T, Tsuboi T, Kitaguchi T
  TITLE     RGB-color intensiometric indicators visualize spatiotemporal 
            dynamics of ATP in single cells.
  JOURNAL   Angew Chem Int Ed Engl. 2018 Jun 27. doi: 10.1002/anie.201804304.
  PUBMED    29952110
REFERENCE   2  (bases 1 to 6552)
  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"; journalName: "Angew Chem 
            Int Ed Engl. 2018 Jun 27. doi: 10.1002/anie.201804304."
COMMENT     Sequence Label: MaLionB
FEATURES             Location/Qualifiers
     source          1..6552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(85..104)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        276..655
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        656..859
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     810..830
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     904..923
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        904..922
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1330..1349)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2091..2112
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2184..2201)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2190..2414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2460..2888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2547..2566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2757..2778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2897..2917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2902..3231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3082..3217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3144..3163
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3298..4092
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3352..3371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3962..3981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4266..4387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4303..4322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4357..4376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4436..4452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4436..4452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4449..4471)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4460..4476
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4484..4490,4491..4508,4509..4514))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4529..4550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4667..4684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4838..5426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4918..4937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5597..6388,6389..6457))
                     /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     6220..6239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6458..6552,1..10))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
       61 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
      121 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
      181 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      241 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      301 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      361 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      421 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      481 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      541 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      601 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      661 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      721 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      781 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      841 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      901 aattaatacg actcactata gggagaccca agctggctag cgtttaaacg ggccctctag
      961 actcgagcgg ccgccactgt gctggatatc tgcagaattc caccacactg gactagtgga
     1021 tccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcca ggtcgagctg
     1081 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggtgaggg cgatgccacc
     1141 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1201 accctcgtga ccaccctgtc acacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1261 aagcagcacg acttcttcaa gtccgccatg cccggaggct acatccagga gcgcaccatc
     1321 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1381 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1441 cacaagctgg agtacactct ccgctttatg aagaccgtga aggtgaacat caccaccccc
     1501 gacggccccg tgtacgacgc cgacatcgag atggtgagcg tgcgggccga gagcggcgac
     1561 ctgggcatcc tgcccggcca catccccacc aaggcccccc tgaagatcgg cgccgtgcgg
     1621 ctgaagaagg acggccagac cgagtacgtg gccgtgagcg gcggcaacgt ggaggtgcgg
     1681 cccgaccagg tgaccatcac cgcccaggcc gccgagaccg ccgagggcat cgacaaggag
     1741 cgggccgagg ccgcccggca gcgggcccag gagcggctga acagccagag cgacgacacc
     1801 gacatccggc gggccgagct ggccctgcag cgggccctga accggctgga cgtggccgag
     1861 atgaagactg aattctcacc atataactgt tatatcaagg ccgacaagca gaagaacggc
     1921 atcaaggcga acttcaagat ccgccacaac atcgagggcg gcggcgtgca gctcgcctac
     1981 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2041 ctgagcgtgc agtccatact ttcgaaagac cccaacgaga agcgcgatca catggtcctg
     2101 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaaag
     2161 cttaagttta aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2221 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2281 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2341 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2401 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2461 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2521 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2581 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2641 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2701 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2761 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2821 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2881 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3061 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3121 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3181 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3241 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3301 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3361 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3421 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3481 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3541 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3601 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3661 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3721 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3781 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3841 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3901 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3961 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4021 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4081 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4141 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4201 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4261 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4321 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4381 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4441 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4501 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4561 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4621 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4681 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4741 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4801 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4861 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4921 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4981 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5041 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5101 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5161 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5221 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5281 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5341 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5401 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5461 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5521 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5581 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5641 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5701 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5761 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5821 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5881 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5941 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6001 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6061 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6121 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6181 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6241 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6301 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6361 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6421 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6481 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6541 ataaacaaat ag
//
