LOCUS       Exported                6549 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Red ATP sensor.
ACCESSION   .
VERSION     .
KEYWORDS    MaLionR
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6549
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        162..541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        542..745
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     696..716
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     790..809
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        790..808
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1326..1345
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2067..2084)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2073..2297
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2343..2771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2430..2449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2640..2661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2780..2800)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2785..3114
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2965..3100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3027..3046
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3181..3975
                     /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(3235..3254)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3845..3864
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4149..4270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4186..4205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4240..4259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4319..4335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4319..4335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4332..4354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4343..4359
                     /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(4367..4397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4412..4433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4550..4567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4721..5309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4801..4820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5480..6340)
                     /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     6103..6122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6341..6445)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6520..6539)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
       61 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      121 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      181 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      241 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      301 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      361 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      421 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      481 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      541 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      601 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      661 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      721 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
      781 tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt taaacgggcc
      841 ctctagactc gagcggccgc cactgtgctg gatatctgca gaattccacc acactggact
      901 agtggatcca tgctggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      961 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1021 gagggcgagg gccgccccta cgaggccttt cagaccgcta agctgaaggt gaccaagggt
     1081 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1141 tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc cgagggcttc
     1201 aggtgggagc gcgtgatgaa cttcgaggac ggcggcatta ttcacgttaa ccaggactcc
     1261 tccctgcagg acggcgtatt catctacaag gtgaagctgc gcggcaccaa cttccccccc
     1321 gacggccccg taatgcagaa gaagaccatg ggctgggagc cttctcgctc tatgaagacc
     1381 gtgaaggtga acatcaccac ccccgacggc cccgtgtacg acgccgacat cgagatggtg
     1441 agcgtgcggg ccgagagcgg cgacctgggc atcctgcccg gccacatccc caccaaggcc
     1501 cccctgaaga tcggcgccgt gcggctgaag aaggacggcc agaccgagta cgtggccgtg
     1561 agcggcggca acgtggaggt gcggcccgac caggtgacca tcaccgccca ggccgccgag
     1621 accgccgagg gcatcgacaa ggagcgggcc gaggccgccc ggcagcgggc ccaggagcgg
     1681 ctgaacagcc agagcgacga caccgacatc cggcgggccg agctggccct gcagcgggcc
     1741 ctgaaccggc tggacgtggc cgagatgaag cctgaagagg tttccgagcg gatgtacccc
     1801 gaggacggcg ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac
     1861 tacgccgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     1921 tacatcgtcg acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa
     1981 cagtgcgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     2041 aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2101 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2161 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2221 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2281 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2341 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2401 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2461 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2521 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2581 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2641 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2701 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2761 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2821 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2881 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2941 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3001 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3061 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3121 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3181 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3241 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3301 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3361 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3421 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3481 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3541 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3601 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3661 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3721 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3781 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3841 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3901 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3961 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4021 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4081 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4141 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4201 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4261 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4321 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4381 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4561 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5161 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5221 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5281 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5341 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5401 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5461 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5521 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5581 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5641 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5701 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5761 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5821 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5881 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5941 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6001 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6061 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6121 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6181 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6241 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6301 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6361 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6421 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6481 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     6541 atagttaag
//
