LOCUS       sequence-198420-        5857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses membrane displayed ATP sensor in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5857)
  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 5857)
  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: GfaABC1D-pm-iATPSnFR1.0
FEATURES             Location/Qualifiers
     source          1..5857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   153..282
                     /label=AAV2 ITR
     intron          1133..1265
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1248..1267
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1310..1328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     1334..1393
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1942..1963
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2066..2087)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2570..2599
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2603..2749
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     polyA_signal    2776..2897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2787..2806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2841..2860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3059..3188
                     /label=AAV2 ITR
     primer_bind     complement(3207..3224)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3207..3223)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3216..3238)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3365..3820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3452..3471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3662..3683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3846..3950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3951..4019,4020..4811)
                     /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"
     rep_origin      4982..5570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5471..5490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaga tttaattaag gccttaatta ggctgcgcgc tcgctcgctc actgaggccg
      181 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
      241 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
      301 cgccatgcta cttatctacg tagccatgct ctaggaagat ctaacatatc ctggtgtgga
      361 gtaggggacg ctgctctgac agaggctcgg gggcctgagc tggctctgtg agctggggag
      421 gaggcagaca gccaggcctt gtctgcaagc agacctggca gcattgggct ggccgccccc
      481 cagggcctcc tcttcatgcc cagtgaatga ctcaccttgg cacagacaca atgttcgggg
      541 tgggcacagt gcctgcttcc cgccgcaccc cagcccccct caaatgcctt ccgagaagcc
      601 cattgagcag ggggcttgca ttgcacccca gcctgacagc ctggcatctt gggataaaag
      661 cagcacagcc ccctaggggc tgcccttgct gtgtggcgcc accggcggtg gagaacaagg
      721 ctctattcag cctgtgccca ggaaagggga tcaggggatg cccaggcatg gacagtgggt
      781 ggcagggggg gagaggaggg ctgtctgctt cccagaagtc caaggacaca aatgggtgag
      841 gggagagctc tccccatagc tgggctgcgg cccaacccca ccccctcagg ctatgccagg
      901 gggtgttgcc aggggcaccc gggcatcgcc agtctagccc actccttcat aaagccctcg
      961 catcccagga gcgagcagag ccagagcagg ttggagagga gacgcatcac ctccgctgct
     1021 cgcaagcttt attgcggtag tttatcacag ttaaattgct aacgcagtca gtgcttctga
     1081 cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat
     1141 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     1201 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     1261 cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc
     1321 actataggct agcatggaga cagacacact cctgctatgg gtactgctgc tctgggttcc
     1381 aggttccact ggtgacagat ctatgaagac tattcacgtg agtgtcgtaa ctcccgacgg
     1441 gcctgtatat gaggatgacg ttgaaatggt gagcgtcaaa gcaaaaagtg gcgagctcgg
     1501 tattctccca ggccacattc ccttgaaagc tcccctagag atcagtgccg cacgcctgaa
     1561 gaaagggggc aaaacacagt atatcgctgt gtcaggcggc aacctcgaag tgcggcctga
     1621 caaggtgacc atctatgccc aggcagccga gagggctgag gatatcgatg tcctgcgcgc
     1681 caaggccgcc aaggagagag ccgagcgccg actccaatca caggtcctga gccacaacgt
     1741 ctatatcacc gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     1801 cgtggaggac ggcagcatgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1861 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgtgc tgagcaaaga
     1921 ccctaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1981 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatgagca agggcgagga
     2041 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2101 gttcagcgtg cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     2161 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2221 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2281 cgccatgccc gaaggctacg tccaggagcg caccatcagc ttcaaggacg acggcaccta
     2341 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2401 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactttgg
     2461 gttgcacgac atcgatttta agcgcgccga gctcgccctt aagcgcgcaa tgaataggct
     2521 ctcagttgcc gaaatgaagg gagggaccgg cggtagcctg caggtcgacg aacaaaaact
     2581 catctcagaa gaggatctga atgctgtggg ccaggacacg caggaggtca tcgtggtgcc
     2641 acactccttg ccctttaagg tggtggtgat ctcagccatc ctggccctgg tggtgctcac
     2701 catcatctcc cttatcatcc tcatcatgct ttggcagaag aagccacgtt aggcggccgc
     2761 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2821 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2881 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2941 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3001 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     3061 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3121 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3181 gcgcgcagcc ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa
     3241 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3301 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3361 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3421 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3481 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3541 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3601 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3661 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3721 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3781 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     3841 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3901 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3961 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4021 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4081 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4141 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4201 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4261 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4321 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4381 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4441 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4501 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4561 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4621 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4681 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4741 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4801 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4861 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4921 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4981 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5041 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5101 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5161 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5221 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5281 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5341 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5401 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5461 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5521 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5581 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5641 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5701 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5761 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5821 ttcccgactg gaaagcgggc agtgagcgca acgcaat
//
