LOCUS       sequence-198421-        5578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses cytoplasmic  ATP sensor in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5578)
  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 5578)
  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-cyto-iATPSnFR1.0
FEATURES             Location/Qualifiers
     source          1..5578
                     /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"
     primer_bind     1873..1894
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1997..2018)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2497..2618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2508..2527)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2562..2581
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2780..2909
                     /label=AAV2 ITR
     primer_bind     complement(2928..2945)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2928..2944)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2937..2959)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3086..3541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3173..3192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3383..3404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3567..3671
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3672..3740,3741..4532)
                     /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      4703..5291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5192..5211
                     /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 agcatgaaga ctattcacgt gagtgtcgta actcccgacg ggcctgtata
     1381 tgaggatgac gttgaaatgg tgagcgtcaa agcaaaaagt ggcgagctcg gtattctccc
     1441 aggccacatt cccttgaaag ctcccctaga gatcagtgcc gcacgcctga agaaaggggg
     1501 caaaacacag tatatcgctg tgtcaggcgg caacctcgaa gtgcggcctg acaaggtgac
     1561 catctatgcc caggcagccg agagggctga ggatatcgat gtcctgcgcg ccaaggccgc
     1621 caaggagaga gccgagcgcc gactccaatc acaggtcctg agccacaacg tctatatcac
     1681 cgccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acgtggagga
     1741 cggcagcatg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1801 gctgctgccc gacaaccact acctgagcac ccagtccgtg ctgagcaaag accctaacga
     1861 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1921 ggacgagctg tacaagggcg gtaccggagg gagcatgagc aagggcgagg agctgttcac
     1981 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2041 gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac
     2101 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2161 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2221 cgaaggctac gtccaggagc gcaccatcag cttcaaggac gacggcacct acaagacccg
     2281 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2341 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactttg ggttgcacga
     2401 catcgatttt aagcgcgccg agctcgccct taagcgcgca atgaataggc tctcagttgc
     2461 cgaaatgaag taggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     2521 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2581 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2641 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     2701 aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     2761 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2821 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2881 gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt
     2941 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3001 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3061 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3241 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3301 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3361 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3421 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3481 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3541 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3601 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3661 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3721 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3781 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3841 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3901 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3961 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4021 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4081 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4141 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4201 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4261 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4321 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4381 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4441 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4501 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4561 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4621 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4681 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4741 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4801 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4861 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4921 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4981 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5041 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5101 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5161 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5221 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5281 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5341 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5401 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5461 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5521 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaat
//
