LOCUS       sequence-361797-        4490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self complementary AAV vector for high-throughput in vivo 
            measurement of enhancer activity by massively parallel reporter 
            assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4490)
  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 4490)
  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: scAAV-Mlc2v-MPRA
FEATURES             Location/Qualifiers
     source          1..4490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      436..445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             442..1152
                     /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(587..605)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     855..874
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1242..1259)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1248..1455
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1543..1672
                     /label=AAV2 ITR
     primer_bind     complement(1691..1708)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1691..1707)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1700..1722)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1849..2304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1936..1955)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2146..2167
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2330..2434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2435..2503,2504..3295)
                     /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(2653..2672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3466..4054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3955..3974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4342..4363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4378..4383,4384..4401,4402..4408)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4416..4432
                     /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)."
     primer_bind     4421..4443
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4440..4456
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4440..4456
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
       61 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagagag ggagtggtta
      121 agctagagcg gacttcggtc cgctttttgg taccttagac aatggcagga cccagagcac
      181 agagcatcgt tcccaggcca ggccccagcc actgtctctt taaccttgaa ggcatttttg
      241 ggtctcacgt gtccacccag gcgggtgtcg gactttgaac ggctcttact tcagaagaac
      301 ggcatggggt gggggggctt aggtggcctc tgcctcacct acaactgcca aaagtggtca
      361 tggggttatt tttaacccca gggaagaggt atttattgtt ccacagcagg ggccggccag
      421 caggctcctt gaattgccac catggtgagc aagggcgagg aggataacat ggccatcatc
      481 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagtttgag
      541 attgaaggcg aaggcgaggg cagaccctac gagggaacac agaccgccaa gctgaaggtg
      601 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
      661 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
      721 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      781 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
      841 ttcccctccg acggccccgt aatgcagaag aagacaatgg gctgggaggc ctcctccgag
      901 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
      961 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     1021 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     1081 accatcgtgg aacagtacga acgcgccgag ggtcgacact ccaccggcgg catggacgag
     1141 ctgtacaagt agctgcggcc gcatcagagc tagcttcagc atcgatactt gaggcgcgcc
     1201 tagccatctc gaggacgggg tgaactacgc ctgaggatct gcctcgactg tgccttctag
     1261 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1321 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1381 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1441 caggcatgct ggggactcga gttaagggcg aattcccgat aaggatcttc ctagagcatg
     1501 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     1561 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     1621 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt
     1681 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     1741 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     1801 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     1861 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1921 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1981 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2041 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2101 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2161 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2221 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2281 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2341 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2401 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2461 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2521 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2581 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2641 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2701 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2761 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2821 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2881 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2941 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3001 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3061 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3121 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3181 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3241 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3301 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3361 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3421 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3481 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3541 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3601 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3661 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3721 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3781 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3841 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3901 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3961 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4021 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4081 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4141 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4201 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     4261 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4321 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     4381 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     4441 aggaaacagc tatgaccatg attacgccag atttaattaa gggatctggg
//
