LOCUS       sequence-413336-        6731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-responsive control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6731)
  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 6731)
  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).cpSFGFP.HaloTag
FEATURES             Location/Qualifiers
     source          1..6731
                     /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     1508..1529
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1632..1653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2967..2984
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3007..3040)
                     /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(3103..3136)
                     /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(3174..3193)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3255..3277)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3556..3833)
                     /label=chicken beta-actin promoter
     repeat_region   4144..4273
                     /label=AAV2 ITR
     rep_origin      4345..4800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4432..4451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4642..4663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4826..4930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4931..4999,5000..5791)
                     /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(5149..5168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5962..6550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6451..6470
                     /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 gatctagcca caacgtctat atcatggccg
     1321 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacgtg gaggacggca
     1381 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1441 tgcccgacaa ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc
     1501 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1561 agctgtacaa gggcggtacc ggagggagca tgagcaaggg cgaggagctg ttcaccgggg
     1621 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg
     1681 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     1741 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1801 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1861 gctacgtcca ggagcgcacc atcagcttca aggacgacgg cacctacaag acccgcgccg
     1921 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1981 aggaggacgg caacatcctg gggcataagc tggagtacaa ctttaacctg cagggcggag
     2041 gctcgggcgg aggtgggtcg ggtggcggcg gatcaatggc cgagatcggc accggcttcc
     2101 ccttcgaccc ccactacgtg gaggtgctgg gcgagcgcat gcactacgtt gacgtgggcc
     2161 cccgcgacgg cacccccgtg ctgttcctgc acggcaaccc caccagcagc tacgtgtggc
     2221 gcaacatcat cccccacgtg gcccccaccc accgcactat cgcccccgac ctgatcggca
     2281 tgggcaagag cgacaagccc gacctgggct acttcttcga cgaccacgtg cgcttcatgg
     2341 acgccttcat cgaggccctg ggcctggagg aggtggtgct ggtgatccac gactggggca
     2401 gcgccctggg cttccactgg gccaagcgca accccgagcg cgtgaagggc atcgccttca
     2461 tggagttcat ccgccccatc cccacctggg acgagtggcc cgagttcgcc cgcgagacct
     2521 tccaggcctt ccgcaccacc gacgtgggcc gcaagctgat catcgaccag aacgtgttca
     2581 tcgagggcac cctgcccatg ggcgtggtgc gccccctgac cgaggtggag atggaccact
     2641 accgcgagcc cttcctgaac cccgtggacc gcgagcccct gtggcgcttc cccaacgagc
     2701 tgcccatcgc cggcgagccc gccaacatcg tggccctggt ggaggagtac atggactggc
     2761 tgcaccagag ccccgtgccc aagctgctgt tctggggcac ccccggcgtg ctgatccccc
     2821 ccgccgaggc cgcccgcctg gccaagagcc tgcccaacgt taaggccgtg gacatcggcc
     2881 ccggcctgaa cctgctccag gaggacaacc ccgacctgat cggcagcgag atcgcccgct
     2941 ggctgagcac cctcgagatc agcggccatc accatcacca ccattaagaa ttcgaagctt
     3001 gctagtataa cttcgtatag catacattat acgaagttat gggtcgatgg tgatgcttgg
     3061 caattcgggt cgatggtgaa gcattggtct tcctattttg atataacttc gtataggata
     3121 ctttatacga agttatgggg atgatccaat tctttgccaa aatgatgaga cagcacaata
     3181 accagcacgt tgcccaggag ctgtaggaaa aagaagaagg catgaacatg gttagcagag
     3241 gctctagagc cgccggtcac acgccagaag ccgaaccccg ccctgccccg tcccccccga
     3301 aggcagccgt ccccccgcgg acagccccga gccgctcccc ccgcacaaag ggccctcccg
     3361 gagcccctca aggctttcac gcagccacag aaaagaaaca agccgtcatt aaaccaagcg
     3421 ctaattacag cccggaggag aagggccgtc ccgcccgctc acctgtggga gtaacgcggt
     3481 cagtcagagc cggggcgggc ggcgcgaggc ggcggcggag cggggcacgg ggcgaaggca
     3541 gcgcgcagcg actcccgccc gccgcgcgct tcgcttttta tagggccgcc gccgccgccg
     3601 cctcgccata aaaggaaact ttcggagcgc gccgctctga ttggctgccg ccgcacctct
     3661 ccgcctcgcc ccgccccscc ccycsccccs ccccsccccs ccyggcgcgc gccccccccc
     3721 cccccccgcc cccatcgctg cacaaaataa ttaaaaaata aataaataca aaattggggg
     3781 tggggagggg ggggagatgg ggagagtgaa gcagaacgtg gggctcacct cgaccatggt
     3841 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     3901 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     3961 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     4021 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     4081 tggtactctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     4141 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4201 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4261 agcgagcgcg cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4321 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4381 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4441 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4501 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4561 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4621 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4681 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4741 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4801 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4861 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4921 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4981 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5041 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5101 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5161 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5221 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5281 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5341 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5401 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5461 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5521 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5581 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5641 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5701 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5761 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5941 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6001 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6061 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6121 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6181 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6241 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6301 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6361 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6421 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6481 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6541 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6601 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6661 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6721 ggaagcggaa g
//
