LOCUS       sequence-330892-        8529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the ATP sensor GRAB_ATP1.0 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8529)
  AUTHORS   Wu Z, He K, Chen Y, Li H, Pan S, Li B, Liu T, Xi F, Deng F, Wang H, 
            Du J, Jing M, Li Y
  TITLE     A sensitive GRAB sensor for detecting extracellular ATP in vitro and
            in vivo.
  JOURNAL   Neuron. 2021 Dec 17. pii: S0896-6273(21)00988-0. doi: 
            10.1016/j.neuron.2021.11.027.
  PUBMED    34942116
REFERENCE   2  (bases 1 to 8529)
  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: "Neuron. 
            2021 Dec 17. pii: S0896-6273(21)00988-0. doi: 
            10.1016/j.neuron.2021.11.027."
COMMENT     Sequence Label: pDisplay-GRAB_ATP1.0-IRES-mCherry-CAAX
FEATURES             Location/Qualifiers
     source          1..8529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     737..796
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1822..1843
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1949..1970)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2210..2229)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(2727..3301,3302..3304,3305..3313)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2895..2912)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3121..3140
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3314..4021
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3459..3477)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3727..3746
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4079..4108
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4112..4258
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(4280..4297)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4286..4510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4642..5230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4722..4741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5559..5606
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5631..5650
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5838..6632)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTYGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5949..5968)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6559..6578
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(6731..6750)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        6829..7020
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6977..6997
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(7080..7871,7872..7940))
                     /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     7703..7722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7941..8045)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8072..8527
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8159..8178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8369..8390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccctgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg
      721 cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
      781 tccaggttcc actggtgaca caagtttgtg cgaaaaagtt ggcaccaccg aggtgctgtg
      841 gccggctgtc cccaacggga cggacgctgc cttcctggcc ggtccgggtt cgtcctgggg
      901 gaacagcacg gtcgcctcca ctgccgccgt ctcctcgtcg ttcaaatgcg ccttgaccaa
      961 gacgggcttc cagttttact acctgccggc tgtctacatc ttggtattca tcatcggctt
     1021 cctgggcaac agcgtggcca tctggatgtt cgtcttccac atgaagccct ggagcggcat
     1081 ctccgtgtac atgttcaatt tggctctggc cgacttcttg tacgtgctga ctctgccagc
     1141 cctgatcttc tactacttca ataaaacaga ctggatcttc ggggatgcca tgtgtaaact
     1201 gcagaggttc atctttcatg tgaacctcta tggcagcatc ttgtttctga catgcatcag
     1261 tgcccaccgg tacagcggtg tggtgtaccc cctcaagtcc ctgggccggc tcaaaaagaa
     1321 gaatgcgatc tgtatcagcg tgctggtgtg gctcattgtg gtggtggcga tctcccccat
     1381 cctcttctac tcaggtaccg gggtccgcaa aaacaaaacc atcacctgtt acgacaccac
     1441 ctcagacgag tacctgcgaa gttatttcat ctacagcatg tgcacgaccg tggccatgtt
     1501 ctgtgtcccc ttggtgctga ttctgggctg ttacggatta attgtgagag ctttgattta
     1561 ccccgcgccg gccgggccgc gcgacaccga cgcgctggac ctggaggagg gaggaaacgt
     1621 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttccaca tccgccacaa
     1681 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1741 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     1801 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1861 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gaaagggcga
     1921 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1981 caagttcagc gtgtccggcg agggtgaggg cgatgccacc gagggcaagc tgaccctgaa
     2041 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2101 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2161 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     2221 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2281 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacac
     2341 cggagcagca gcacgctggc gcgggcggtc gatttacctg gtaatcattg tactgactgt
     2401 ttttgctgtg tcttacatcc ctttccatgt gatgaaaacg atgaacttga gggcccggct
     2461 tgattttcag accccagcaa tgtgtgcttt caatgacagg gtttatgcca cgtatcaggt
     2521 gacaagaggt ctagcaagtc tcaacagttg tgtggacccc attctctatt tcttggcggg
     2581 agatactttc agaaggagac tctcccgagc cacaaggaaa gcttctagaa gaagtgaggc
     2641 aaatttgcaa tccaagagtg aagacatgac cctcaatatt ttacctgagt tcaagcagaa
     2701 cggagataca agcctgtaag ctagcgcccc tctccctccc ccccccccta acgttactgg
     2761 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     2821 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     2881 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     2941 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     3001 gaacccccca cctggcgaca ggtgcctctg cggccaacgc cacgtgtata agatacacct
     3061 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     3121 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     3181 atgggatctg atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa
     3241 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     3301 tatggccaca accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     3361 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     3421 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     3481 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     3541 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     3601 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     3661 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     3721 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     3781 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     3841 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     3901 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     3961 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     4021 ggatggtaaa aagaagaaaa agaagtcaaa gacaaagtgt gtaattatgt aagtcgacga
     4081 acaaaaactc atctcagaag aggatctgaa tgctgtgggc caggacacgc aggaggtcat
     4141 cgtggtgcca cactccttgc cctttaaggt ggtggtgatc tcagccatcc tggccctggt
     4201 ggtgctcacc atcatctccc ttatcatcct catcatgctt tggcagaaga agccacgtta
     4261 ggcggccgct cgagatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4321 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4381 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4441 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4501 ggctctatgg cttctgaggc ggaaagaacc agtggcggta atacggttat ccacagaatc
     4561 gggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac gcgacttatc
     4981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5041 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     5401 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     5461 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     5521 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     5581 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     5641 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     5701 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     5761 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     5821 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     5881 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     5941 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     6001 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     6061 gcatcgccat aggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     6121 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     6181 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     6241 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     6301 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     6361 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     6421 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     6481 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     6541 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     6601 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     6661 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     6721 agaggccggg cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa
     6781 tgggcggaac tgggcggagt taggggcgga atgggcggag ttaggggcgg gactatggtt
     6841 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     6901 ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     6961 cctggggact ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag
     7021 gactcttcct ttttcaataa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7081 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7141 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7201 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7261 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7321 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7381 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7441 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7501 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7561 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7621 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7681 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7741 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7801 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7861 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7921 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8041 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     8101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     8161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     8221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     8281 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     8341 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     8401 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     8461 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     8521 acaatttac
//
