LOCUS       sequence-228536-        6099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear localized mRuby2FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  AUTHORS   Economo MN, Viswanathan S, Tasic B, Bas E, Winnubst J, Menon V, 
            Graybuck LT, Nguyen TN, Smith KA, Yao Z, Wang L, Gerfen CR, 
            Chandrashekar J, Zeng H, Looger LL, Svoboda K
  TITLE     Distinct descending motor cortex pathways and their roles in 
            movement.
  JOURNAL   Nature. 2018 Nov;563(7729):79-84. doi: 10.1038/s41586-018-0642-9. 
            Epub 2018 Oct 31.
  PUBMED    30382200
REFERENCE   2  (bases 1 to 6099)
  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"; doi: 
            "10.1038/s41586-018-0642-9"; journalName: "Nature"; date: "2018-11";
            volume: "563"; issue: "7729"; pages: "79-84"
COMMENT     Sequence Label: pAAV- CAG-h2BmRuby2FLAG
FEATURES             Location/Qualifiers
     source          1..6099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     313..335
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     397..416
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             479..856
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             872..895
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             899..922
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             926..949
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1454..1477
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1484..1507
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1520..1543
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1550..1573
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1766..1789
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1793..1816
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1828..1842
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     misc_feature    1861..2449
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1914..1934)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2332..2343)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2492..2613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2503..2522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2557..2576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2775..2904
                     /label=AAV2 ITR
     rep_origin      complement(3141..3729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3221..3240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3900..4691,4692..4760))
                     /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     4523..4542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4761..4865)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4891..5346)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5028..5049)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5240..5259
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5418..5547
                     /label=AAV2 ITR
     promoter        join(5858..6099,1..34)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
       61 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
      121 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
      181 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
      241 tttgtgcggg gggagcggct cggggctgtc cgcgggggga cggctgcctt cgggggggac
      301 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
      361 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
      421 ctcatcattt tggcaaagaa ttggatccga attcactagt ctcgaggata tcgccaccat
      481 gccagagcca gcgaagtctg ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa
      541 ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc aaggagagct attccatcta
      601 tgtgtacaag gttctgaagc aggtccaccc tgacaccggc atttcgtcca aggccatggg
      661 catcatgaat tcgtttgtga acgacatttt cgagcgcatc gcaggtgagg cttcccgcct
      721 ggcgcattac aacaagcgct cgaccatcac ctccagggag atccagacgg ccgtgcgcct
      781 gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa
      841 gtacaccagc gctaaggatc caggtaccat ggactacaag gacgacgacg acaaaggtga
      901 ctacaaagat gatgacgata aaggcgacta taaggacgat gacgacaagg gcggaaactc
      961 actgatcaag gaaaacatgc ggatgaaggt ggtgatggag ggctccgtga atggtcacca
     1021 gttcaagtgc accggagagg gagagggaaa cccgtacatg ggaactcaga ccatgcgcat
     1081 taaggtcatc gaaggaggtc cgctgccgtt cgctttcgat atcctggcca cttcgttcgg
     1141 aggagggtcg cgcacgttca tcaagtaccc gaagggaatc ccggacttct ttaagcagtc
     1201 attcccggaa ggattcactt gggaacgggt gacccggtat gaagatggag gtgtggtgac
     1261 tgtcatgcaa gatacttcgc tggaggatgg gtgcctcgtg taccacgtcc aagtccgcgg
     1321 agtgaatttc ccgtccaacg gaccagtgat gcagaaaaag acgaagggtt gggaacctaa
     1381 tactgaaatg atgtaccccg cagacggagg gctgaggggc tacacccaca tggcgctgaa
     1441 ggtcgacgga ggagattaca aggatgacga cgataagcaa caagattaca aagacgatga
     1501 tgacaagggc cagcagggcg actacaagga cgacgacgac aagcagcagg actacaaaga
     1561 tgacgatgat aaaggaggag gacatctgtc ctgttcgttc gtgaccacct acagatcaaa
     1621 gaaaaccgtg ggaaacatca agatgccggg cattcatgcc gtcgaccacc gcctggagcg
     1681 gctcgaagaa tcagacaatg agatgttcgt cgtgcaaaga gaacatgccg tggccaagtt
     1741 cgcgggactg ggaggcggtg gaggcgatta caaagacgat gatgacaagg gtgactataa
     1801 agacgacgat gacaaagggg attaaaggat gatgatgata agtagtaaaa gcttatcgat
     1861 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1921 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     1981 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2041 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2101 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2161 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2221 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     2281 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     2341 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     2401 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     2461 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2521 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2581 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2641 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     2701 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     2761 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2821 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2881 ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2941 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3001 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3061 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3121 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3181 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3241 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3301 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3361 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3421 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3481 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3541 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3601 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3661 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3721 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3781 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3841 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3901 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3961 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4021 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4081 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4141 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4201 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4261 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4321 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4381 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4441 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4501 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4561 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4621 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4681 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4741 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4801 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4861 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     4921 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4981 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5041 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5101 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     5161 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5221 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5281 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5341 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5401 cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     5461 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     5521 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
     5581 ctacgtagcc atgctctagg aagagtacca ttgacgtcaa taatgacgta tgttcccata
     5641 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     5701 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     5761 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5821 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     5881 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     5941 ttattttgtg cagcgatggg ggcggggggg gggggggcgc gsgccaggcg gggcggggcg
     6001 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
     6061 gctccgaaag tttcctttta tggcgaggcg gcggcggcg
//
