LOCUS       sequence-335423-        6833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MeCP2 Knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6833)
  AUTHORS   Gulmez Karaca K, Brito DVC, Zeuch B, Oliveira AMM
  TITLE     Adult hippocampal MeCP2 preserves the genomic responsiveness to 
            learning required for long-term memory formation.
  JOURNAL   Neurobiol Learn Mem. 2018 Mar;149:84-97. doi: 
            10.1016/j.nlm.2018.02.010. Epub 2018 Feb 10.
  PUBMED    29438740
REFERENCE   2  (bases 1 to 6833)
  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: "Neurobiol 
            Learn Mem."; date: "2018-03"; volume: "149:84-97. doi"; pages: " 
            10.1016/j.nlm.2018.02.010. Epub 2018 Feb 10"
COMMENT     Sequence Label: rAAV-U6-MeCP2shRNA-CamKIIa-mCherry
FEATURES             Location/Qualifiers
     source          1..6833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        112..130
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     112..129
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   151..280
                     /label=AAV2 ITR
     promoter        445..693
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     445..465
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     616..635
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             2045..2755
                     /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(2190..2208)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2458..2477
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2803..3391
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2856..2876)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3274..3285)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3418..3435)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3424..3648
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3858..4187)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3872..4007
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4172..4192
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4433..5021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4513..4532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5192..5983,5984..6052))
                     /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     5815..5834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6053..6157)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6225..6243
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6281..6303)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6403..6422
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6636..6655
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatgatagg gtctgcttca gtaagccaga tgctacacaa ttaggcttgt acatattgtc
       61 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
      121 acactataga atacacggaa ttaattctag ctgcgcgctc gctcgctcac tgaggccgcc
      181 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
      241 cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg
      301 ccatgctact tatctacgta gccatgctct aggtaccctc gaggaggaga agcatgaatt
      361 ccccagtgga aagacgcgca ggcaaaacgc accacgtgac ggagcgtgac cgcgcgccga
      421 gcgcgcgcca aggtcgggca ggaagagggc ctatttccca tgattccttc atatttgcat
      481 atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata
      541 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      601 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      661 ggtttatata tcttgtggaa aggacgcggg atcccgtcag aagaccagga tctccattca
      721 agagatggag atcctggtct tctgactttt ttggaaagat ctaagcttgg cactggccgt
      781 actagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag ggtcacatca
      841 gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat gccttggggg
      901 catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca atgagtttag
      961 gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg gtgagtccag
     1021 ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga agcacaagca
     1081 gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg aggatgggtg
     1141 gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga cagatagggg
     1201 tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac ttcttctcac
     1261 gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg gaaagaaaca
     1321 aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa agaaaccatt
     1381 acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt aaaccttaga
     1441 acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga tgaagttgcc
     1501 agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc cactagccgt
     1561 taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa aagggcccta
     1621 tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag tttgttcgca
     1681 tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg
     1741 gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa
     1801 agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc
     1861 ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca
     1921 cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag
     1981 gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatcag gtggcagcgg
     2041 tggcatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     2101 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     2161 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     2221 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     2281 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     2341 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     2401 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     2461 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     2521 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     2581 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     2641 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     2701 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtagggtgg
     2761 cagcggtggc actcgagtaa gctatcaagc tagcttatcg ataatcaacc tctggattac
     2821 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2881 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2941 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3001 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3061 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3121 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3181 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     3241 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3301 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3361 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgactcgc tgatcagcct
     3421 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3481 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3541 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3601 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3661 aaagaaccag ctggggctcg actagagcat ggctacgtag ataagtagca tggcgggtta
     3721 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3781 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga
     3841 gcgagcgcgc agagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     3901 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     3961 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     4021 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     4081 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     4141 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgc
     4201 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4261 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4321 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4381 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4441 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4501 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4561 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4621 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4681 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4741 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4801 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4861 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4921 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4981 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5041 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5101 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5161 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5221 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5281 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5341 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5401 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5461 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5521 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5581 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5641 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5701 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5761 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5821 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5881 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5941 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6001 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6061 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6181 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6241 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6301 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6361 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6421 tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     6481 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     6541 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     6601 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     6661 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg
     6721 ctagcgatga ccctgctgat tggttcgctg accatttccg ggtgcgggac ggcgttacca
     6781 gaaactcaga aggttcgtcc aaccaaaccg actctgacgg cagtttacga gag
//
