LOCUS       sequence-196778-        6240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cytoplasmic ATP sensor for mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6240)
  AUTHORS   Lobas MA, Tao R, Nagai J, Kronschlager MT, Borden PM, Marvin JS, 
            Looger LL, Khakh BS
  TITLE     A genetically encoded single-wavelength sensor for imaging cytosolic
            and cell surface ATP.
  JOURNAL   Nat Commun. 2019 Feb 12;10(1):711. doi: 10.1038/s41467-019-08441-5.
  PUBMED    30755613
REFERENCE   2  (bases 1 to 6240)
  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: "Nat 
            Commun."; date: "2019-02-12"; volume: "10(1):711. doi"; pages: " 
            10.1038/s41467-019-08441-5"
COMMENT     Sequence Label: cyto-iATPSnFR1.0
FEATURES             Location/Qualifiers
     source          1..6240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             731..757
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1442..1463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1928..1957
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1982..1999)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1988..2212
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2344..2932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2424..2443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3261..3308
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3333..3352
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3540..4334)
                     /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
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3651..3670)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4261..4280
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4433..4452)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        4531..4722
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4679..4699
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4782..5573,5574..5642))
                     /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     5405..5424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5643..5747)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5774..6229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5861..5880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6071..6092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag aactctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgaac tcggatccac tagtaacggc cgccagtgtg ctggaattcg gcttggggat
      721 atccaccatg tatccatacg atgttccaga ttatgctggg gcccagccgg ccagatctat
      781 gaagactatt cacgtgagtg tcgtaactcc cgacgggcct gtatatgagg atgacgttga
      841 aatggtgagc gtcaaagcaa aaagtggcga gctcggtatt ctcccaggcc acattccctt
      901 gaaagctccc ctagagatca gtgccgcacg cctgaagaaa gggggcaaaa cacagtatat
      961 cgctgtgtca ggcggcaacc tcgaagtgcg gcctgacaag gtgaccatct atgcccaggc
     1021 agccgagagg gctgaggata tcgatgtcct gcgcgccaag gccgccaagg agagagccga
     1081 gcgccgactc caatcacagg tcctgagcca caacgtctat atcaccgccg acaagcagaa
     1141 gaacggcatc aaggcgaact tcaagatccg ccacaacgtg gaggacggca gcatgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgtgctgag caaagaccct aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gggcggtacc ggagggagca tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1441 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga
     1501 gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1561 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1621 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1681 ggagcgcacc atcagcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt
     1741 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1801 caacatcctg gggcacaagc tggagtacaa ctttgggttg cacgacatcg attttaagcg
     1861 cgccgagctc gcccttaagc gcgcaatgaa taggctctca gttgccgaaa tgaagctgca
     1921 ggtcgacgaa caaaaactca tctcagaaga ggatctgaat taggcggccg ctcgagatca
     1981 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2041 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2101 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2161 gaggattggg aagacaatag caggcgtgct ggggatgcgg tgggctctat ggcttctgag
     2221 gcggaaagaa ccagtggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2761 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3061 taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac
     3121 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
     3181 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
     3241 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt
     3301 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     3361 ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     3421 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     3481 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     3541 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     3601 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     3661 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     3721 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     3781 acgagatcct cgccgtcggg cttgctcgcc ttgagcctgg cgaacagttc ggctggcgcg
     3841 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     3901 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     3961 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     4021 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     4081 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     4141 gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     4201 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     4261 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     4321 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc
     4381 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg ggcctcggcc
     4441 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     4501 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     4561 ttgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     4621 taattgagat gcttgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4681 cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat
     4741 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4801 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4861 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4921 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4981 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5041 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5101 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5161 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5221 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5281 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5341 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5401 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5461 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5521 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5581 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5641 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5701 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5761 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5821 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6121 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6181 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgcgcgcgtt
//
