LOCUS       sequence-384461-        5080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV9 based MPRA assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5080)
  AUTHORS   Akerberg BN, Gu F, VanDusen NJ, Zhang X, Dong R, Li K, Zhang B, Zhou
            B, Sethi I, Ma Q, Wasson L, Wen T, Liu J, Dong K, Conlon FL, Zhou J,
            Yuan GC, Zhou P, Pu WT
  TITLE     A reference map of murine cardiac transcription factor chromatin 
            occupancy identifies dynamic and conserved enhancers.
  JOURNAL   Nat Commun. 2019 Oct 28;10(1):4907. doi: 10.1038/s41467-019-12812-3.
  PUBMED    31659164
REFERENCE   2  (bases 1 to 5080)
  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: "Nat 
            Commun."; date: "2019-10-28"; volume: "10(1):4907. doi"; pages: " 
            10.1038/s41467-019-12812-3"
COMMENT     Sequence Label: aav-hsp68-mCherry-MCS-pA
FEATURES             Location/Qualifiers
     source          1..5080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..62
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     54..71
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     55..71
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   90..219
                     /label=AAV2 ITR
     polyA_signal    307..514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     503..520
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(610..1320)
                     /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(888..907)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1157..1175
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2386..2402)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2386..2402)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2399..2421)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2410..2426
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2434..2440,2441..2458,2459..2464))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2479..2500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2788..3376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2868..2887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3547..4338,4339..4407))
                     /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     4170..4189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4408..4512)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4538..4993)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4675..4696)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4887..4906
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
       61 cgacggccag tgaattaggt taattaaggc tgcgcgctcg ctcgctcact gaggccgccc
      121 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
      181 gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc
      241 catgctactt atctacgtag ccatgctcta ggaagatcct tatcgggaat tcgcccttaa
      301 ctcgagtccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
      361 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
      421 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
      481 caaacaacag atggctggca actagaaggc acagtcgagg cagatcctca ggcgtagttc
      541 accccgtcct cgagatggct aggcgcgcct caagtatcga tgctgaagct agctctgatg
      601 cggccgcagc tacttgtaca gctcgtccat gccgccggtg gagtgtcgac cctcggcgcg
      661 ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt
      721 gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc
      781 agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc
      841 gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccattgtct tcttctgcat
      901 tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc
      961 gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac
     1021 gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg
     1081 gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa
     1141 gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgtgttccct cgtagggtct
     1201 gccctcgcct tcgccttcaa tctcaaactc gtggccgttc acggagccct ccatgtgcac
     1261 cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct tgctcaccat
     1321 ggcgccgcgc tctgcttctg gaaggctgcg ctccgcggcg tggatgctcc ggggaaagtt
     1381 gcgctccgcg gcagggatgc tcctgggaag gttgcgctcc gcggcaggga tgctctgggg
     1441 aaggctggtc ctggccgagg atcgggaacg cgccgctcgc tctgcttctc ttgtcttcgc
     1501 ttgtctctgg atggaaccag atttggttct gagtagctgt cagcgtctgg tgacctgctc
     1561 gccgtcctgc gcctttaagg agtcttcacc ggccccgccc actctccgct gggccaatca
     1621 gcgagccgga ggaggccttg gggccaggaa tcttccagca gtttcgcgtc tggtggagct
     1681 tccccgcctc ccttgagtaa tcggagttgt gggttccgcc cttgtccaga actctccaga
     1741 ggtttctggg gttcactgga gagtacggat tcctgagggg gagggtgtgg ggaagtgctg
     1801 gtgctactag tgacactgtt gctatggcga cgcattacta aggcctgtgt ggaatggaca
     1861 agaaagatca cctctagctc ggtgttgtgt acagtttgtt gtgatttgtg gggtttcgcc
     1921 aactcgcaca gttctgaata tgggggttaa aggctaaaac ttaagggcta aaacttctcc
     1981 ccgccaagtt taggagaccc agggagatgc ctgggggcgt gtccggtgac gtgatcctct
     2041 ccaatcgcgt tacaatggca gtgctgcctc tgacctcatg gactaattta ggaactagag
     2101 gctctgtccc agcacaggct caaagttgcc gggaggggcg gggtgggggg tgggggggac
     2161 cccggctgct cagtttggat gttcctggag ctcggtacca aaaagcggac cgaagtccgc
     2221 tctagcttaa ccactccctc tctgcgcgct cgctcgctcg ctcactgagg ccgggcgacc
     2281 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2341 agagggagtg gcccagatcc cttaattaaa tctggcgtaa tcatggtcat agctgtttcc
     2401 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2461 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2521 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4501 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     4561 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4621 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4681 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4741 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4801 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4861 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4921 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4981 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5041 cgggcctctt cgctattacg ccagctggcg aaagggggat
//
