LOCUS       sequence-433791-        6652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of iATPSnFR2, high affinity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6652)
  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 6652)
  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.CAG.(cyto).iATPSnFR2.S29W.A95K.HaloTag
FEATURES             Location/Qualifiers
     source          1..6652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(278..866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(358..377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1037..1828,1829..1897))
                     /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     1660..1679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1898..2002)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2028..2483)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2186)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2377..2396
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2555..2684
                     /label=AAV2 ITR
     primer_bind     3553..3575
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3637..3656
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3690..3699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4247..4268
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4371..4392)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    5799..6387
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5852..5872)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6270..6281)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6430..6551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6441..6460)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6495..6514
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgact
      121 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      181 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      241 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      301 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      361 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      421 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      481 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      541 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      601 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      661 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      721 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      781 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      841 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      901 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      961 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1021 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1081 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1141 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1201 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1261 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1321 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1381 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1441 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1501 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1561 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1621 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1681 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1741 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1801 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1861 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1921 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1981 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     2041 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2101 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2161 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2221 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2281 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2341 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2401 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2461 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
     2521 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
     2581 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     2641 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
     2701 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcgtaccat tgacgtcaat
     2761 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     2821 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     2881 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     2941 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     3001 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     3061 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
     3121 cgcgccgggg gggggggggg gggggggggg gsggggsggg gsgrggsggr grggkgcggc
     3181 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     3241 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
     3301 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
     3361 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
     3421 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
     3481 tttgtgcggg gggagcggct cggggctgtc cgcgggggga cggctgcctt cgggggggac
     3541 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
     3601 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     3661 ctcatcattt tggcaaagaa ttggatcccg ccaccatggg cagatctatg aagactattc
     3721 acgtgagtgt cgtaactccc gacgggcctg tatatgagga tgacgttgaa atggtgagcg
     3781 tcaaagcaaa atggggcgag ctcggtattc tcccaggcca cattcccttg aaagctcccc
     3841 tagagatcag tgccgcacgc ctgaagaaag ggggcaaaac acagtatatc gctgtgtcag
     3901 gcggcaacct cgaagtgcgg cctgacaagg tgaccatcta tgcccaggca gccgagaggg
     3961 ctgaggatat cgatgtcctg cgcgccaaga aagccaagga gagagccgag cgccgactcc
     4021 aatcacaggt cctggtcggg aacgtctata tcaccgccga caagcagaag aacggcatca
     4081 aggcgaactt caagatccgc cacaacgtgg aggacggcag catgcagctc gccgaccact
     4141 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4201 gcacccagtc cgtgctgagc aaagacccta acgagaagcg cgatcacatg gtcctgctgg
     4261 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg
     4321 gagggagcat gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4381 tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca
     4441 ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     4501 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     4561 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     4621 tcagcttcaa ggacgacggc acctacaaga cccgcgccga ggtgaagttc gagggcgaca
     4681 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     4741 ggcacaagct ggagtacaac tttatttgtg tggacatcga ttttaagcgc gccgagctcg
     4801 cccttaagcg cgcaatgaat aggctctcag ttgccgaaat gaagctgcag ggcggaggct
     4861 cgggcggagg tgggtcgggt ggcggcggat caatggccga gatcggcacc ggcttcccct
     4921 tcgaccccca ctacgtggag gtgctgggcg agcgcatgca ctacgttgac gtgggccccc
     4981 gcgacggcac ccccgtgctg ttcctgcacg gcaaccccac cagcagctac gtgtggcgca
     5041 acatcatccc ccacgtggcc cccacccacc gcactatcgc ccccgacctg atcggcatgg
     5101 gcaagagcga caagcccgac ctgggctact tcttcgacga ccacgtgcgc ttcatggacg
     5161 ccttcatcga ggccctgggc ctggaggagg tggtgctggt gatccacgac tggggcagcg
     5221 ccctgggctt ccactgggcc aagcgcaacc ccgagcgcgt gaagggcatc gccttcatgg
     5281 agttcatccg ccccatcccc acctgggacg agtggcccga gttcgcccgc gagaccttcc
     5341 aggccttccg caccaccgac gtgggccgca agctgatcat cgaccagaac gtgttcatcg
     5401 agggcaccct gcccatgggc gtggtgcgcc ccctgaccga ggtggagatg gaccactacc
     5461 gcgagccctt cctgaacccc gtggaccgcg agcccctgtg gcgcttcccc aacgagctgc
     5521 ccatcgccgg cgagcccgcc aacatcgtgg ccctggtgga ggagtacatg gactggctgc
     5581 accagagccc cgtgcccaag ctgctgttct ggggcacccc cggcgtgctg atcccccccg
     5641 ccgaggccgc ccgcctggcc aagagcctgc ccaacgttaa ggccgtggac atcggccccg
     5701 gcctgaacct gctccaggag gacaaccccg acctgatcgg cagcgagatc gcccgctggc
     5761 tgagcaccct cgagatcagc ggctagaagc ttatcgataa tcaacctctg gattacaaaa
     5821 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5881 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5941 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     6001 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     6061 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     6121 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     6181 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     6241 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     6301 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6361 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag
     6421 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     6481 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     6541 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     6601 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aa
//
