LOCUS       sequence-413355-        6177 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 6177)
  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 6177)
  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).cpSFGFP.HaloTag
FEATURES             Location/Qualifiers
     source          1..6177
                     /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     1668..1689
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1792..1813)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3127..3144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3179..3767
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3232..3252)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3650..3661)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3795..3916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3806..3825)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3860..3879
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4078..4207
                     /label=AAV2 ITR
     rep_origin      complement(4444..5032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4524..4543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5203..5994,5995..6063))
                     /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(6064..6168)
                     /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 gatctagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1501 aaggcgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac
     1561 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1621 agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1681 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc
     1741 ggagggagca tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1801 ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc
     1861 accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1921 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     1981 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2041 atcagcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac
     2101 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2161 gggcataagc tggagtacaa ctttaacctg cagggcggag gctcgggcgg aggtgggtcg
     2221 ggtggcggcg gatcaatggc cgagatcggc accggcttcc ccttcgaccc ccactacgtg
     2281 gaggtgctgg gcgagcgcat gcactacgtt gacgtgggcc cccgcgacgg cacccccgtg
     2341 ctgttcctgc acggcaaccc caccagcagc tacgtgtggc gcaacatcat cccccacgtg
     2401 gcccccaccc accgcactat cgcccccgac ctgatcggca tgggcaagag cgacaagccc
     2461 gacctgggct acttcttcga cgaccacgtg cgcttcatgg acgccttcat cgaggccctg
     2521 ggcctggagg aggtggtgct ggtgatccac gactggggca gcgccctggg cttccactgg
     2581 gccaagcgca accccgagcg cgtgaagggc atcgccttca tggagttcat ccgccccatc
     2641 cccacctggg acgagtggcc cgagttcgcc cgcgagacct tccaggcctt ccgcaccacc
     2701 gacgtgggcc gcaagctgat catcgaccag aacgtgttca tcgagggcac cctgcccatg
     2761 ggcgtggtgc gccccctgac cgaggtggag atggaccact accgcgagcc cttcctgaac
     2821 cccgtggacc gcgagcccct gtggcgcttc cccaacgagc tgcccatcgc cggcgagccc
     2881 gccaacatcg tggccctggt ggaggagtac atggactggc tgcaccagag ccccgtgccc
     2941 aagctgctgt tctggggcac ccccggcgtg ctgatccccc ccgccgaggc cgcccgcctg
     3001 gccaagagcc tgcccaacgt taaggccgtg gacatcggcc ccggcctgaa cctgctccag
     3061 gaggacaacc ccgacctgat cggcagcgag atcgcccgct ggctgagcac cctcgagatc
     3121 agcggccatc accatcacca ccattaagaa ttcgaagctt ttcgaattag cttatcataa
     3181 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3241 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3301 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3361 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3421 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3481 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3541 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3601 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3661 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3721 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc ggaggccgct
     3781 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3841 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3901 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3961 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     4021 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     4081 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4141 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4201 cgcgcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4261 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4321 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4381 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4441 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4501 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4561 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4621 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4681 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4741 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4801 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4861 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4921 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4981 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5041 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5101 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5161 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5221 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5281 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5341 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5401 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5461 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5521 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5581 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5641 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5701 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5761 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5821 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5881 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5941 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6001 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6061 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6121 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttcc
//
