LOCUS       sequence-207273-        7074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid to target the ecAT3.10 extracellular 
            ATP sensor to the cell surface..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7074)
  AUTHORS   Conley JM, Radhakrishnan S, Valentino SA, Tantama M
  TITLE     Imaging extracellular ATP with a genetically-encoded, ratiometric 
            fluorescent sensor.
  JOURNAL   PLoS One. 2017 Nov 9;12(11):e0187481. doi: 
            10.1371/journal.pone.0187481. eCollection 2017.
  PUBMED    29121644
REFERENCE   2  (bases 1 to 7074)
  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: "PLoS One.";
            date: "2017-11-9"; volume: "12(11):e0187481. doi"; pages: " 
            10.1371/journal.pone.0187481. eCollection 2017"
COMMENT     Sequence Label: ecAT3.10
FEATURES             Location/Qualifiers
     source          1..7074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        26..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     560..580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     654..673
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        654..672
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     753..812
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             join(819..821,822..824,825..1337)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             join(1917..1919,1920..1922,1923..2633)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2640..2669
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2673..2819
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2841..2858)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2847..3071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3203..3791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3283..3302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4120..4167
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4192..4211
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4399..5193)
                     /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(4510..4529)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5120..5139
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5292..5311)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5390..5581
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5538..5558
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5641..6432,6433..6501))
                     /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     6264..6283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6502..6606)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6633..7061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6720..6739)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6930..6951
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacgcttaca atttacgcgc gcgttgacat tgattattga ctagttatta atagtaatca
       61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      421 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      541 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      601 agcagaactc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
      661 actcactata gggagaccca agcttggtac cgaactcgga tccactagta acggccgcca
      721 gtgtgctgga attcggcttg gggatatcca ccatggagac agacacactc ctgctatggg
      781 tactgctgct ctgggttcca ggttccactg gtgacagaat ggtgagcaag ggcgaggagc
      841 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaggt
      901 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      961 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctggg
     1021 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1081 ccatgcccga aggctacgtc caggagcgta ccatcttctt caaggacgac ggcaactaca
     1141 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1201 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacatca
     1261 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc cacttcaaga
     1321 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1381 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     1441 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1501 ccggtatcga tatgaaaacg atccacgtga gcgtcactac tcctgatggc ccggtgtacg
     1561 aagacgatgt tgaaatggtc agcgtcaaag cgaaaagcgg cgagctcggc attttgccgg
     1621 ggcacattcc gcttaaggcc ccgctcgaga tcagcgcggc ccggctgaaa aaaggcggca
     1681 aaacgcaata cattgccgtc agcggcggca atttggaagt ccgcccggac aaagtgacga
     1741 ttaacgctca agctgctgaa cgggcggagg acattgacgt cctccgcgcc aaagcggcga
     1801 aagagcgggc ggagcgccgc ctgcaaagcc agcaggacga catcgacttc aaacgggccg
     1861 aactggcgtt aaaacgcgcc atgaaccgtt tgagcgttgc cgaaatgaag gaattcatgg
     1921 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1981 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2041 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2101 tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     2161 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2221 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2281 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2341 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     2401 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     2461 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2521 tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2581 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggtcgacg
     2641 aacaaaaact catctcagaa gaggatctga atgctgtggg ccaggacacg caggaggtca
     2701 tcgtggtgcc acactccttg ccctttaagg tggtggtgat ctcagccatc ctggccctgg
     2761 tggtgctcac catcatctcc cttatcatcc tcatcatgct ttggcagaag aagccacgtt
     2821 aggcggccgc tcgagatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2881 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2941 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3001 ggggcaggac agcaaggggg aggattggga agacaatagc aggcgtgctg gggatgcggt
     3061 gggctctatg gcttctgagg cggaaagaac cagtggcggt aatacggtta tccacagaat
     3121 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3181 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3241 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3301 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3361 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3421 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3481 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3541 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3601 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3661 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3721 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3781 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3901 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg
     3961 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     4021 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     4081 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt
     4141 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
     4201 tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc
     4261 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     4321 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     4381 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     4441 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     4501 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     4561 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     4621 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc ttgctcgcct tgagcctggc
     4681 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4741 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4801 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4861 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4921 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     4981 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     5041 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     5101 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     5161 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     5221 atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5281 cagaggccgg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     5341 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5401 tgctgactaa ttgagatgct tgctttgcat acttctgcct gctggggagc ctggggactt
     5461 tccacacctg gttgctgact aattgagatg cttgctttgc atacttctgc ctgctgggga
     5521 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     5581 ggactcttcc tttttcaata aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6601 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     6661 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     6721 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     6781 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6841 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     6901 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6961 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     7021 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tatt
//
