LOCUS       sequence-329817-        5782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mRuby in CaMKII+ cells with low miRNA-128 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5782)
  AUTHORS   Keaveney MK, Rahsepar B, Tseng HA, Fernandez FR, Mount RA, Ta T, 
            White JA, Berg J, Han X
  TITLE     CaMKIIalpha-Positive Interneurons Identified via a microRNA-Based 
            Viral Gene Targeting Strategy.
  JOURNAL   J Neurosci. 2020 Dec 9;40(50):9576-9588. doi: 
            10.1523/JNEUROSCI.2570-19.2020. Epub 2020 Nov 6.
  PUBMED    33158963
REFERENCE   2  (bases 1 to 5782)
  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: 
            "10.1523/JNEUROSCI.2570-19.2020"; journalName: "J Neurosci"; date: 
            "2020-12-9- 9"; volume: "40"; issue: "50"; pages: "9576-9588"
COMMENT     Sequence Label: pAAV-CaMKII-mRuby2-4x128-mAGNET
FEATURES             Location/Qualifiers
     source          1..5782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     regulatory      1503..1512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1509..2219
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    2369..2957
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2422..2442)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2840..2851)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2960..2976)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2961..2977)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3006..3127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3017..3036)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3071..3090
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3506..4094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3586..3605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4265..5056,5057..5125))
                     /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     4888..4907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5126..5230)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5256..5711)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5393..5414)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5605..5624
                     /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 gaattccatt atggccttag gtaggtcact tcatctccat ggggttcttc
      241 ttctgatttt ctagaaaatg agatgggggt gcagagagct tcctcagtga cctgcccagg
      301 gtcacatcag aaatgtcaga gctagaactt gaactcagat tactaatctt aaattccatg
      361 ccttgggggc atgcaagtac gatatacaga aggagtgaac tcattagggc agatgaccaa
      421 tgagtttagg aaagaagagt ccagggcagg gtacatctac accacccgcc cagccctggg
      481 tgagtccagc cacgttcacc tcattatagt tgcctctctc cagtcctacc ttgacgggaa
      541 gcacaagcag aaactgggac aggagcccca ggagaccaaa tcttcatggt ccctctggga
      601 ggatgggtgg ggagagctgt ggcagaggcc tcaggagggg ccctgctgct cagtggtgac
      661 agataggggt gagaaagcag acagagtcat tccgtcagca ttctgggtct gtttggtact
      721 tcttctcacg ctaaggtggc ggtgtgatat gcacaatggc taaaaagcag ggagagctgg
      781 aaagaaacaa ggacagagac agaggccaag tcaaccagac caattcccag aggaagcaaa
      841 gaaaccatta cagagactac aagggggaag ggaaggagag atgaattagc ttcccctgta
      901 aaccttagaa cccagctgtt gccagggcaa cggggcaata cctgtctctt cagaggagat
      961 gaagttgcca gggtaactac atcctgtctt tctcaaggac catcccagaa tgtggcaccc
     1021 actagccgtt accatagcaa ctgcctcttt gccccactta atcccatccc gtctgttaaa
     1081 agggccctat agttggaggt gggggaggta ggaagagcga tgatcacttg tggactaagt
     1141 ttgttcgcat ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact
     1201 tgctcctggg cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc
     1261 aggttttaaa gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca
     1321 gggcgtttcg gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt
     1381 gagcagccac agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt
     1441 tgcactcagg agcacgggca ggcgagtggc ccctagttct gggggcagcg gggcgcgccg
     1501 ccgccaccat ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg
     1561 tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc
     1621 cgtacatggg aactcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg
     1681 cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga
     1741 aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta
     1801 cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct
     1861 gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc
     1921 agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc
     1981 tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt
     2041 tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg
     2101 ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac
     2161 gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt
     2221 aaactagtgc ggccttaaag agaccggttc actgtgacag taaaagagac cggttcactg
     2281 tgagaatgaa agagaccggt tcactgtgat cggaaaagag accggttcac tgtgagcggc
     2341 cgccctgcag gcatgcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2401 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2461 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2521 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2581 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2641 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2701 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2761 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2821 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2881 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2941 ttgggccgcc tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga
     3001 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3061 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3121 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3181 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     3241 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     3301 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgactcg ctgcgctcgg
     3361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5221 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     5281 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5341 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5401 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5461 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5521 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5581 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5641 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5701 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5761 ggcctcttcg ctattacgcc ag
//
