LOCUS       sequence-413356-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of iATPSnFR2, low affinity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6557)
  AUTHORS   Marvin JS, Kokotos AC, Kumar M, Pulido C, Tkachuk AN, Yao JS, Brown 
            TA, Ryan TA
  TITLE     iATPSnFR2: A high-dynamic-range fluorescent sensor for monitoring 
            intracellular ATP.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 May 21;121(21):e2314604121. doi: 
            10.1073/pnas.2314604121. Epub 2024 May 15.
  PUBMED    38748581
REFERENCE   2  (bases 1 to 6557)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-05-21"; volume: "121(21):e2314604121. 
            doi"; pages: " 10.1073/pnas.2314604121. Epub 2024 May 15"
COMMENT     Sequence Label: pAAV.GFAP.(cyto).iATPSnFR2.A95A.A119L.HaloTag
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(17..472)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(154..175)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     366..385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   544..673
                     /label=AAV2 ITR
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1995..2016
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2119..2140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    3559..4147
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3612..3632)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4030..4041)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4175..4296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4186..4205)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4240..4259
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4458..4587
                     /label=AAV2 ITR
     rep_origin      complement(4824..5412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4904..4923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5583..6374,6375..6443))
                     /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"
     promoter        complement(6444..6548)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
       61 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
      121 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
      181 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
      241 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
      301 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
      361 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
      421 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
      481 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      541 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
      601 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      661 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      721 tctaggaaga tcgtaggtta attaataaca tatcctggtg tggagtaggg gacgctgctc
      781 tgacagaggc tcgggggcct gagctggctc tgtgagctgg ggaggaggca gacagccagg
      841 cctttgtctg caagcagacc tggcagcatt gggctggccg ccccccaggg cctcctcttc
      901 atgcccagtg aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg
      961 cttcccgccg caccccagcc cccctcaaat gccttccgag aagcccattg agcagggggc
     1021 ttgcattgca ccccagcctg acagcctggc atcttgggat aaaagcagca cagcccccta
     1081 ggggctgccc ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta ttcagcctgt
     1141 gcccaggaaa ggggatcagg ggatgcccag gcatggacag tgggtggcag ggggggagag
     1201 gagggctgtc tgcttcccag aagtccaagg acacaaatgg gtgaggggag agctctcccc
     1261 atagctgggc tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg
     1321 cacccgggca tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag
     1381 cagagccaga gcaggttgga gaggagacgc atcacctccg ctgctcgcgg ggatcccgcc
     1441 accatgggca gatctatgaa gactattcac gtgagtgtcg taactcccga cgggcctgta
     1501 tatgaggatg acgttgaaat ggtgagcgtc aaagcaaaaa gtggcgagct cggtattctc
     1561 ccaggccaca ttcccttgaa agctccccta gagatcagtg ccgcacgcct gaagaaaggg
     1621 ggcaaaacac agtatatcgc tgtgtcaggc ggcaacctcg aagtgcggcc tgacaaggtg
     1681 accatctatg cccaggcagc cgagagggct gaggatatcg atgtcctgcg cgccaaggcc
     1741 gccaaggaga gagccgagcg ccgactccaa tcacaggtcc tggtcgggaa cgtctatatc
     1801 accgccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca caacgtggag
     1861 gacggcagca tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1921 gtgctgctgc ccgacaacca ctacctgagc acccagtccg tgctgagcaa agaccctaac
     1981 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2041 atggacgagc tgtacaaggg cggtaccgga gggagcatga gcaagggcga ggagctgttc
     2101 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2161 gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc
     2221 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2281 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2341 cccgaaggct acgtccagga gcgcaccatc agcttcaagg acgacggcac ctacaagacc
     2401 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2461 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt tatttgtgtg
     2521 gacatcgatt ttaagcgcgc cgagctcttg cttaagcgcg caatgaatag gctctcagtt
     2581 gccgaaatga agctgcaggg cggaggctcg ggcggaggtg ggtcgggtgg cggcggatca
     2641 atggccgaga tcggcaccgg cttccccttc gacccccact acgtggaggt gctgggcgag
     2701 cgcatgcact acgttgacgt gggcccccgc gacggcaccc ccgtgctgtt cctgcacggc
     2761 aaccccacca gcagctacgt gtggcgcaac atcatccccc acgtggcccc cacccaccgc
     2821 actatcgccc ccgacctgat cggcatgggc aagagcgaca agcccgacct gggctacttc
     2881 ttcgacgacc acgtgcgctt catggacgcc ttcatcgagg ccctgggcct ggaggaggtg
     2941 gtgctggtga tccacgactg gggcagcgcc ctgggcttcc actgggccaa gcgcaacccc
     3001 gagcgcgtga agggcatcgc cttcatggag ttcatccgcc ccatccccac ctgggacgag
     3061 tggcccgagt tcgcccgcga gaccttccag gccttccgca ccaccgacgt gggccgcaag
     3121 ctgatcatcg accagaacgt gttcatcgag ggcaccctgc ccatgggcgt ggtgcgcccc
     3181 ctgaccgagg tggagatgga ccactaccgc gagcccttcc tgaaccccgt ggaccgcgag
     3241 cccctgtggc gcttccccaa cgagctgccc atcgccggcg agcccgccaa catcgtggcc
     3301 ctggtggagg agtacatgga ctggctgcac cagagccccg tgcccaagct gctgttctgg
     3361 ggcacccccg gcgtgctgat cccccccgcc gaggccgccc gcctggccaa gagcctgccc
     3421 aacgttaagg ccgtggacat cggccccggc ctgaacctgc tccaggagga caaccccgac
     3481 ctgatcggca gcgagatcgc ccgctggctg agcaccctcg agatcagcgg ctagaagctt
     3541 ttcgaattag cttatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3601 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3661 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3721 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3781 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3841 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3901 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3961 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4021 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4081 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4141 tccccgcatc ggaggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     4201 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4261 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     4321 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     4381 tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg
     4441 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4501 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4561 ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg cgcggggaga
     4621 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5221 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5881 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6181 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6421 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6481 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6541 gttccgcgca catttcc
//
