LOCUS       sequence-355955-        6823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a control shRNA with no known targets in the mouse genome 
            under control of the U6 promoter and mCherry under control of the 
            CaMKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  AUTHORS   Hagenston AM, Yan J, Bas-Orth C, Tan Y, Sekler I, Bading H
  TITLE     Disrupted expression of mitochondrial NCLX sensitizes neuroglial 
            networks to excitotoxic stimuli and renders synaptic activity toxic.
  JOURNAL   J Biol Chem. 2021 Dec 20:101508. doi: 10.1016/j.jbc.2021.101508.
  PUBMED    34942149
REFERENCE   2  (bases 1 to 6823)
  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: "J Biol 
            Chem. 2021 Dec 20:101508. doi: 10.1016/j.jbc.2021.101508."
COMMENT     Sequence Label: pAAV-shCTRL
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     promoter        298..546
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     298..318
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     469..488
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             1888..2598
                     /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(2033..2051)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2301..2320
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2646..3234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2699..2719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3117..3128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3261..3278)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3267..3491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3701..4030)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3715..3850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4015..4035
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4276..4864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4356..4375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5035..5826,5827..5895))
                     /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     5658..5677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5896..6000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6068..6086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6124..6146)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6246..6265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6479..6498
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6788..6806
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6788..6805
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc ctcgaggagg agaagcatga attccccagt ggaaagacgc gcaggcaaaa
      241 cgcaccacgt gacggagcgt gaccgcgcgc cgagcgcgcg ccaaggtcgg gcaggaagag
      301 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
      361 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
      421 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
      481 cttaccgtaa cttgaaagta tttcgatttc ttgggtttat atatcttgtg gaaaggacgc
      541 gggatctccg tgccaagacg ggtagtcatt caagagatga ctacccgtct tggcactttt
      601 tggaagagct tggcactggc cgtactagaa aatgagatgg gggtgcagag agcttcctca
      661 gtgacctgcc cagggtcaca tcagaaatgt cagagctaga acttgaactc agattactaa
      721 tcttaaattc catgccttgg gggcatgcaa gtacgatata cagaaggagt gaactcatta
      781 gggcagatga ccaatgagtt taggaaagaa gagtccaggg cagggtacat ctacaccacc
      841 cgcccagccc tgggtgagtc cagccacgtt cacctcatta tagttgcctc tctccagtcc
      901 taccttgacg ggaagcacaa gcagaaactg ggacaggagc cccaggagac caaatcttca
      961 tggtccctct gggaggatgg gtggggagag ctgtggcaga ggcctcagga ggggccctgc
     1021 tgctcagtgg tgacagatag gggtgagaaa gcagacagag tcattccgtc agcattctgg
     1081 gtctgtttgg tacttcttct cacgctaagg tggcggtgtg atatgcacaa tggctaaaaa
     1141 gcagggagag ctggaaagaa acaaggacag agacagaggc caagtcaacc agaccaattc
     1201 ccagaggaag caaagaaacc attacagaga ctacaagggg gaagggaagg agagatgaat
     1261 tagcttcccc tgtaaacctt agaacccagc tgttgccagg gcaacggggc aatacctgtc
     1321 tcttcagagg agatgaagtt gccagggtaa ctacatcctg tctttctcaa ggaccatccc
     1381 agaatgtggc acccactagc cgttaccata gcaactgcct ctttgcccca cttaatccca
     1441 tcccgtctgt taaaagggcc ctatagttgg aggtggggga ggtaggaaga gcgatgatca
     1501 cttgtggact aagtttgttc gcatcccctt ctccaacccc ctcagtacat caccctgggg
     1561 gaacagggtc cacttgctcc tgggcccaca cagtcctgca gtattgtgta tataaggcca
     1621 gggcaaagag gagcaggttt taaagtgaaa ggcaggcagg tgttggggag gcagttaccg
     1681 gggcaacggg aacagggcgt ttcggaggtg gttgccatgg ggacctggat gctgacgaag
     1741 gctcgcgagg ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct cgtcagtcaa
     1801 gccggttctc cgtttgcact caggagcacg ggcaggcgag tggcccctag ttctgggggc
     1861 agcgggggat caggtggcag cggtggcatg gtgagcaagg gcgaggagga taacatggcc
     1921 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1981 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2041 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     2101 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2161 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2221 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2281 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2341 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     2401 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     2461 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     2521 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2581 gacgagctgt acaagtaggg tggcagcggt ggcactcgag taagctatca agctagctta
     2641 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3181 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3241 accgtcgact cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3301 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3361 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3421 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3481 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgactagag catggctacg
     3541 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     3601 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3661 gcccgggcgg cctcagtgag cgagcgagcg cgcagagctt tttgcaaaag cctaggcctc
     3721 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     3781 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     3841 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     3901 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     3961 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     4021 actgacacac attccacagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4081 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4141 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4201 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4261 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4321 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4381 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4441 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4501 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4561 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4621 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4681 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4741 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4801 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4861 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4921 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4981 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5041 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5101 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5161 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5221 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5281 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5341 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5401 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5461 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5521 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5581 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5641 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5701 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5761 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5821 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5881 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5941 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6001 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6061 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6121 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6181 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6241 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct cccttatgcg
     6301 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6361 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6421 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6481 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6541 tgcgtccggc gtagaggatc tggctagcga tgaccctgct gattggttcg ctgaccattt
     6601 ccgggtgcgg gacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga
     6661 cggcagttta cgagagagat gatagggtct gcttcagtaa gccagatgct acacaattag
     6721 gcttgtacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     6781 acatacgatt taggtgacac tatagaatac acggaattaa ttc
//
