LOCUS       sequence-196779-        6405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  membrane displayed ATP sensor for mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6405)
  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 6405)
  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: pm-iATPSnFR1.1
FEATURES             Location/Qualifiers
     source          1..6405
                     /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"
     sig_peptide     728..787
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1336..1357
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1460..1481)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1946..1975
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1979..2125
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2147..2164)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2153..2377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2509..3097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2589..2608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3426..3473
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3498..3517
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3705..4499)
                     /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(3816..3835)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4426..4445
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4598..4617)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        4696..4887
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4844..4864
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4947..5738,5739..5807))
                     /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     5570..5589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5808..5912)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5939..6394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6026..6045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6236..6257
                     /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 gagacagaca cactcctgct atgggtactg ctgctctggg ttccaggttc
      781 cactggtgac agatctatga agactattca cgtgagtgtc gtaactcccg acgggcctgt
      841 atatgaggat gacgttgaaa tggtgagcgt caaagcaaaa agtggcgagc tcggtattct
      901 cccaggccac attcccttga aagctcccct agagatcagt gccgcacgcc tgaagaaagg
      961 gggcaaaaca cagtatatcg ctgtgtcagg cggcaacctc gaagtgcggc ctgacaaggt
     1021 gaccatctat gcccaggcag ccgagagggc tgaggatatc gatgtcctgc gcgccaagaa
     1081 agccaaggag agagccgagc gccgactcca atcacaggtc ctgagccaca acgtctatat
     1141 caccgccgac aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacgtgga
     1201 ggacggcagc atgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1261 cgtgctgctg cccgacaacc actacctgag cacccagtcc gtgctgagca aagaccctaa
     1321 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gtcgccggga tcactctcgg
     1381 catggacgag ctgtacaagg gcggtaccgg agggagcatg agcaagggcg aggagctgtt
     1441 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1501 cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
     1561 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1621 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1681 gcccgaaggc tacgtccagg agcgcaccat cagcttcaag gacgacggca cctacaagac
     1741 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1801 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact ttgggttgca
     1861 cgacatcgat tttaagcgcg ccgagctcag ccttaagcgc gcaatgaata ggctctcagt
     1921 tgccgaaatg aagctgcagg tcgacgaaca aaaactcatc tcagaagagg atctgaatgc
     1981 tgtgggccag gacacgcagg aggtcatcgt ggtgccacac tccttgccct ttaaggtggt
     2041 ggtgatctca gccatcctgg ccctggtggt gctcaccatc atctccctta tcatcctcat
     2101 catgctttgg cagaagaagc cacgttaggc ggccgctcga gatcagcctc gactgtgcct
     2161 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2221 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2281 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2341 aatagcaggc gtgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagt
     2401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2701 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2941 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3121 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3181 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3241 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     3301 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     3361 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     3421 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     3481 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     3541 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     3601 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     3661 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     3721 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     3781 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     3841 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     3901 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     3961 tcgggcttgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     4021 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     4081 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     4141 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     4201 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     4261 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     4321 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     4381 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     4441 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     4501 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
     4561 gcctcctcac tacttctgga atagctcaga ggccgggcct cggcctctgc ataaataaaa
     4621 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     4681 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcttgct ttgcatactt
     4741 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcttg
     4801 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     4861 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaataaatc aatctaaagt
     4921 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4981 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5041 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5101 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5161 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5221 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5281 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5341 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5401 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5461 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5521 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5581 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5641 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5701 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5761 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5821 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5881 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5941 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6001 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6061 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6121 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6181 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6241 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6301 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6361 gcgaatttta acaaaatatt aacgcttaca atttacgcgc gcgtt
//
