LOCUS       sequence-357501-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids for production of AAV-based TFactivity reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  AUTHORS   Abe H, Abe K
  TITLE     PCR-based profiling of transcription factor activity in vivo by a 
            virus-based reporter battery.
  JOURNAL   iScience. 2022 Feb 14;25(3):103927. doi: 10.1016/j.isci.2022.103927.
            eCollection 2022 Mar 18.
  PUBMED    35281741
REFERENCE   2  (bases 1 to 5504)
  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: "iScience.";
            date: "2022-02-14"; volume: "25(3):103927. doi"; pages: " 
            10.1016/j.isci.2022.103927. eCollection 2022 Mar 18"
COMMENT     Sequence Label: pAAV-pgk-DIO-rvCRY-WPRE
FEATURES             Location/Qualifiers
     source          1..5504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    complement(168..201)
                     /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)."
     protein_bind    complement(252..285)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(294..1004)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(572..591)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     841..859
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1001..1010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1017..1050
                     /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)."
     protein_bind    1101..1134
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1159..1747
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1212..1232)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1630..1641)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1779..2255
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1978..1997)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2295..2435
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2295..2424
                     /label=AAV2 ITR
     rep_origin      2510..2965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2597..2616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2807..2828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2982..3001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3101..3123
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3161..3179)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3247..3351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3352..3420,3421..4212)
                     /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(3570..3589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4383..4971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4872..4891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5033..5162
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
       61 tcaaaagcgc acgtctgtcg cgctgttctc ctcttcctca tctccgggcc tttcgacctc
      121 acggtaatca ctagtgaatt cgcggccgcc tgcaggtcga ctccggaata acttcgtata
      181 ggatacttta tacgaagtta tgcagaatgg tagctggatt gtagctgcta ttagcaatat
      241 gaaacctctt aataacttcg tatagcatac attatacgaa gttatggcgc gccttacttg
      301 tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg
      361 atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg
      421 ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc gtagtggccg
      481 ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac
      541 atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag
      601 gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag
      661 tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag
      721 ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc
      781 ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc
      841 ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc
      901 tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac
      961 tccttgatga tggccatgtt atcctcctcg cccttgctca ccatggtggc gctagcataa
     1021 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
     1081 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc
     1141 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1201 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1261 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1321 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     1381 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     1441 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1501 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     1561 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     1621 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     1681 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     1741 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     1801 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     1861 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     1921 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     1981 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     2041 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2101 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2161 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     2221 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     2281 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2341 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2401 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2461 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2521 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2581 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2641 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2701 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2761 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2821 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     2881 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2941 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3001 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3061 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3121 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3181 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3841 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3901 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3961 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4021 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4081 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4141 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4201 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4501 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
     5041 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
     5101 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     5161 ctgcggccgc acgcgtcggg agcatgcgac gtcgggccca atctgcagca cgtgccgcgg
     5221 tggcggccgc ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     5281 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     5341 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     5401 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
     5461 gctcagaggc tgggaagggg tgggtccggg ggcgggctca gggg
//
