LOCUS       sequence-218929-        6503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  negative control for Green ATP sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6503)
  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 6503)
  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: negMaLionG
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        466..669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     714..733
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        714..732
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(823..844)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1084..1103)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1660..1911
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1848..1869
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1945..1962)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1951..2175
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2221..2649
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2308..2327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2518..2539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2658..2678)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2663..2992
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2843..2978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2905..2924
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3059..3853
                     /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(3113..3132)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3723..3742
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4027..4148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4064..4083)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4118..4137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4197..4213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4197..4213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4210..4232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4221..4237
                     /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(4245..4251,4252..4269,4270..4275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4290..4311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4428..4445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4599..5187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4679..4698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5358..6149,6150..6218))
                     /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     5981..6000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6219..6323)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6398..6417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
       61 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
      121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      481 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      661 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
      721 actcactata gggagaccca agctggctag cgtttaaacg ggccctctag actcgagatg
      781 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      841 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      901 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      961 gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag
     1021 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1081 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1141 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1201 ctggagtaca acccgcgtgg aatgaagacc gtgaaggtaa acatcaccac ccccgacggc
     1261 cccgtgtacg acgccgacat cgagatggtg agcgtgcggg ccgagagcgg cgacctgggc
     1321 atcctgcccg gccacatccc caccaaggcc cccctgaaga tcggcgccgt gcggctgaag
     1381 aaggacggcc agaccgagta cgtggccgtg agcggcggca acgtggaggt gcggcccgac
     1441 caggtgacca tcaccgccca ggccgccgag accgccgagg gcatcgacaa ggagcgggcc
     1501 gaggccgccc ggcagcgggc ccaggagcgg ctgaacagcc agagcgacga caccgacatc
     1561 cggcgggccg agctggccct gcagcgggcc ctgaaccggc tggacgtggc cgagatgaag
     1621 gcacctggtg aattcagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1681 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1741 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1801 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1861 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaattcgaa
     1921 gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1981 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2041 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2101 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2161 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2221 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2281 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2341 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2401 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2461 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2521 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2581 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2641 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2701 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2761 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2821 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2881 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     2941 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3001 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3061 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3121 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3181 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3241 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3301 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3361 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3421 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3481 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3541 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     3601 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3661 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3721 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3781 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3841 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3901 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3961 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4021 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4081 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4141 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4201 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4261 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4321 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4381 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4441 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4501 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5281 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5341 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5401 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5461 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5521 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5581 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5641 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5701 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5761 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5821 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5881 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5941 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6001 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6061 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6121 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6181 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6241 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6301 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6361 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     6421 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     6481 aaatttaagc tacaacaagg caa
//
