LOCUS       sequence-218931-        6549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  negative control for Red ATP sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6549)
  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 6549)
  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: negMaLionR
FEATURES             Location/Qualifiers
     source          1..6549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        168..547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        548..751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     702..722
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     796..815
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        796..814
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1332..1351
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2073..2090)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2079..2303
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2349..2777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2436..2455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2646..2667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2786..2806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2791..3120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2971..3106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3033..3052
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3187..3981
                     /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(3241..3260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3851..3870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4155..4276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4192..4211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4246..4265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4325..4341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4325..4341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4338..4360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4349..4365
                     /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(4373..4379,4380..4397,4398..4403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4418..4439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4556..4573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4727..5315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4807..4826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5486..6277,6278..6346))
                     /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     6109..6128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6347..6451)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6526..6545)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
       61 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      121 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      181 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      241 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      361 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      421 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      481 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      541 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      601 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      661 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      721 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      781 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agcgtttaaa
      841 cgggccctct agactcgagc ggccgccact gtgctggata tctgcagaat tccaccacac
      901 tggactagtg gatccatgct ggtgagcaag ggcgaggagg ataacatggc catcatcaag
      961 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     1021 gagggcgagg gcgagggccg cccctacgag gcctttcaga ccgctaagct gaaggtgacc
     1081 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     1141 aaggcctaca ttaagcaccc agccgacatc cccgactact tcaagctgtc cttccccgag
     1201 ggcttcaggt gggagcgcgt gatgaacttc gaggacggcg gcattattca cgttaaccag
     1261 gactcctccc tgcaggacgg cgtattcatc tacaaggtga agctgcgcgg caccaacttc
     1321 ccccccgacg gccccgtaat gcagaagaag accatgggct gggagccttc tcgcgggatg
     1381 aagaccgtga aggtgaacat caccaccccc gacggccccg tgtacgacgc cgacatcgag
     1441 atggtgagcg tgcgggccga gagcggcgac ctgggcatcc tgcccggcca catccccacc
     1501 aaggcccccc tgaagatcgg cgccgtgcgg ctgaagaagg acggccagac cgagtacgtg
     1561 gccgtgagcg gcggcaacgt ggaggtgcgg cccgaccagg tgaccatcac cgcccaggcc
     1621 gccgagaccg ccgagggcat cgacaaggag cgggccgagg ccgcccggca gcgggcccag
     1681 gagcggctga acagccagag cgacgacacc gacatccggc gggccgagct ggccctgcag
     1741 cgggccctga accggctgga cgtggccgag atgaagcctg aaggtgtttc cgagcggatg
     1801 taccccgagg acggcgccct gaagagcgag atcaagaagg ggctgaggct gaaggacggc
     1861 ggccactacg ccgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     1921 ggcgcctaca tcgtcgacat caagttggac atcgtgtccc acaacgagga ctacaccatc
     1981 gtggaacagt gcgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     2041 aagtaaaagc ttaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc
     2101 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2161 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2221 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2281 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2341 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2401 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2461 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2521 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2581 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2641 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2701 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2761 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2821 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2941 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3001 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3061 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3121 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3181 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3241 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3301 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3361 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3421 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3481 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3541 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3601 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3661 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3721 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3781 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3841 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3901 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3961 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4021 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4081 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4141 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4201 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4261 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4321 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4381 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4441 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4501 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4561 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4621 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4681 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4741 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4801 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4861 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4921 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4981 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5041 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5101 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5161 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5221 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5281 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5341 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5401 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5461 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5521 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5581 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5641 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5701 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5761 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5821 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5881 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5941 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6001 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6061 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6121 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6181 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6241 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6301 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6361 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6421 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6481 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     6541 tgccgcata
//
