LOCUS       sequence-413202-        7111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of iATPSnFR2, medium affinity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7111)
  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 7111)
  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.CAGFLEX.(cyto).iATPSnFR2.A95K.HaloTag
FEATURES             Location/Qualifiers
     source          1..7111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1133..1166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1237..1270
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1835..1856
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1959..1980)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3387..3420)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3483..3516)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(3554..3573)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3635..3657)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3936..4213)
                     /label=chicken beta-actin promoter
     repeat_region   4524..4653
                     /label=AAV2 ITR
     rep_origin      4725..5180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4812..4831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5022..5043
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5206..5310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5311..5379,5380..6171)
                     /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     complement(5529..5548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6342..6930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6831..6850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct aattcgaacc tcttcgaggg acataacttc
     1141 gtataatgta tgctatacga agttatgggt cgatggtgag atctggacta gagggtcgat
     1201 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     1261 acgaagttat ggatcccgcc accatgggca gatctatgaa gactattcac gtgagtgtcg
     1321 taactcccga cgggcctgta tatgaggatg acgttgaaat ggtgagcgtc aaagcaaaaa
     1381 gtggcgagct cggtattctc ccaggccaca ttcccttgaa agctccccta gagatcagtg
     1441 ccgcacgcct gaagaaaggg ggcaaaacac agtatatcgc tgtgtcaggc ggcaacctcg
     1501 aagtgcggcc tgacaaggtg accatctatg cccaggcagc cgagagggct gaggatatcg
     1561 atgtcctgcg cgccaagaaa gccaaggaga gagccgagcg ccgactccaa tcacaggtcc
     1621 tggtcgggaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gcgaacttca
     1681 agatccgcca caacgtggag gacggcagca tgcagctcgc cgaccactac cagcagaaca
     1741 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1801 tgctgagcaa agaccctaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1861 ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatga
     1921 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1981 taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc
     2041 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2101 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     2161 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg
     2221 acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2281 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2341 agtacaactt tatttgtgtg gacatcgatt ttaagcgcgc cgagctcgcc cttaagcgcg
     2401 caatgaatag gctctcagtt gccgaaatga agctgcaggg cggaggctcg ggcggaggtg
     2461 ggtcgggtgg cggcggatca atggccgaga tcggcaccgg cttccccttc gacccccact
     2521 acgtggaggt gctgggcgag cgcatgcact acgttgacgt gggcccccgc gacggcaccc
     2581 ccgtgctgtt cctgcacggc aaccccacca gcagctacgt gtggcgcaac atcatccccc
     2641 acgtggcccc cacccaccgc actatcgccc ccgacctgat cggcatgggc aagagcgaca
     2701 agcccgacct gggctacttc ttcgacgacc acgtgcgctt catggacgcc ttcatcgagg
     2761 ccctgggcct ggaggaggtg gtgctggtga tccacgactg gggcagcgcc ctgggcttcc
     2821 actgggccaa gcgcaacccc gagcgcgtga agggcatcgc cttcatggag ttcatccgcc
     2881 ccatccccac ctgggacgag tggcccgagt tcgcccgcga gaccttccag gccttccgca
     2941 ccaccgacgt gggccgcaag ctgatcatcg accagaacgt gttcatcgag ggcaccctgc
     3001 ccatgggcgt ggtgcgcccc ctgaccgagg tggagatgga ccactaccgc gagcccttcc
     3061 tgaaccccgt ggaccgcgag cccctgtggc gcttccccaa cgagctgccc atcgccggcg
     3121 agcccgccaa catcgtggcc ctggtggagg agtacatgga ctggctgcac cagagccccg
     3181 tgcccaagct gctgttctgg ggcacccccg gcgtgctgat cccccccgcc gaggccgccc
     3241 gcctggccaa gagcctgccc aacgttaagg ccgtggacat cggccccggc ctgaacctgc
     3301 tccaggagga caaccccgac ctgatcggca gcgagatcgc ccgctggctg agcaccctcg
     3361 agatcagcgg ctagaagctt gctagtataa cttcgtatag catacattat acgaagttat
     3421 gggtcgatgg tgatgcttgg caattcgggt cgatggtgaa gcattggtct tcctattttg
     3481 atataacttc gtataggata ctttatacga agttatgggg atgatccaat tctttgccaa
     3541 aatgatgaga cagcacaata accagcacgt tgcccaggag ctgtaggaaa aagaagaagg
     3601 catgaacatg gttagcagag gctctagagc cgccggtcac acgccagaag ccgaaccccg
     3661 ccctgccccg tcccccccga aggcagccgt ccccccgcgg acagccccga gccgctcccc
     3721 ccgcacaaag ggccctcccg gagcccctca aggctttcac gcagccacag aaaagaaaca
     3781 agccgtcatt aaaccaagcg ctaattacag cccggaggag aagggccgtc ccgcccgctc
     3841 acctgtggga gtaacgcggt cagtcagagc cggggcgggc ggcgcgaggc ggcggcggag
     3901 cggggcacgg ggcgaaggca gcgcgcagcg actcccgccc gccgcgcgct tcgcttttta
     3961 tagggccgcc gccgccgccg cctcgccata aaaggaaact ttcggagcgc gccgctctga
     4021 ttggctgccg ccgcacctct ccgcctcgcc ccgccccscc ccycgccccs ccccsccccs
     4081 ccyggcgcgc gccccccccc cccccccgcc cccatcgctg cacaaaataa ttaaaaaata
     4141 aataaataca aaattggggg tggggagggg ggggagatgg ggagagtgaa gcagaacgtg
     4201 gggctcacct cgaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     4261 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     4321 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     4381 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     4441 tacgtcatta ttgacgtcaa tggtactctt cctagagcat ggctacgtag ataagtagca
     4501 tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct
     4561 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4621 ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag aggcccgcac
     4681 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4741 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4801 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4861 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4921 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4981 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5041 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5101 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5161 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5221 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5281 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5341 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5401 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5461 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5521 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5581 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5641 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5701 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5761 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5821 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5881 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5941 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6001 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6061 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6121 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6181 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6241 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6301 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6361 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6421 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6481 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6541 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6601 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6661 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6721 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6781 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6841 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6901 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6961 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7021 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7081 cgcagcgagt cagtgagcga ggaagcggaa g
//
