LOCUS       sequence-330042-        4813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nuclear mCherry from CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4813)
  AUTHORS   Maturana CJ, Verpeut JL, Kooshkbaghi M, Engel EA
  TITLE     Novel tool to quantify with single-cell resolution the number of 
            incoming AAV genomes co-expressed in the mouse nervous system.
  JOURNAL   Gene Ther. 2021 Jun 28. pii: 10.1038/s41434-021-00272-8. doi: 
            10.1038/s41434-021-00272-8.
  PUBMED    34176926
REFERENCE   2  (bases 1 to 4813)
  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: "Gene Ther. 
            2021 Jun 28. pii: 10.1038/s41434-021-00272-8. doi: 
            10.1038/s41434-021-00272-8."
COMMENT     Sequence Label: pAAV-CMV-mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..4813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        191..394
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     345..365
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             423..1130
                     /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(568..586)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     836..855
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1143..1169
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1170..1196
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1197..1223
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     misc_feature    1251..1839
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1304..1324)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1722..1733)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1842..1858)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1843..1859)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    1888..2009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1899..1918)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1953..1972
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2171..2300
                     /label=AAV2 ITR
     rep_origin      complement(2537..3125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2617..2636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3296..4087,4088..4156))
                     /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     3919..3938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4157..4261)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4287..4742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4424..4445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4636..4655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      241 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      301 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      361 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      421 gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
      481 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      541 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
      601 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
      661 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
      721 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
      781 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
      841 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
      901 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
      961 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1021 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1081 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag cctcgagctg
     1141 atcctgctgc caaacgcgtt aaactagacc ctgctgccaa acgcgttaaa ctagaccctg
     1201 ctgccaaacg cgttaaacta gactaaagcg gccgcgacaa gcttatcgat aatcaacctc
     1261 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1321 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1381 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     1441 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     1501 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     1561 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     1621 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     1681 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     1741 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     1801 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgac
     1861 ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     1921 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1981 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2041 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2101 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     2161 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2221 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2281 gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2941 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4261 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     4321 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4381 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4441 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4501 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4561 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4621 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4681 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4741 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4801 gctattacgc cag
//
