LOCUS       sequence-128338-        6035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses scramble shRNA Cre-dependently by flex switch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6035)
  AUTHORS   Yu X, Ye Z, Houston CM, Zecharia AY, Ma Y, Zhang Z, Uygun DS, Parker
            S, Vyssotski AL, Yustos R, Franks NP, Brickley SG, Wisden W
  TITLE     Wakefulness Is Governed by GABA and Histamine Cotransmission.
  JOURNAL   Neuron. 2015 Jun 17. pii: S0896-6273(15)00516-4. doi: 
            10.1016/j.neuron.2015.06.003.
  PUBMED    26094607
REFERENCE   2  (bases 1 to 6035)
  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: "Neuron. 
            2015 Jun 17. pii: S0896-6273(15)00516-4. doi: 
            10.1016/j.neuron.2015.06.003."
COMMENT     Sequence Label: pAAV-hysn-flex-dsRed-shscramble
FEATURES             Location/Qualifiers
     source          1..6035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(671..704)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(755..788)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     ncRNA           complement(954..1080)
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           complement(1107..1121)
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           complement(1148..1269)
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     CDS             complement(1319..1996)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MDSSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(1381..1404)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1579..1598)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1796..1816
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1848..1866
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    2020..2053
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2104..2137
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2162..2750
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2215..2235)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2633..2644)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2782..3258
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2981..3000)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3298..3438
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3298..3427
                     /label=AAV2 ITR
     rep_origin      3513..3968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3600..3619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3810..3831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3985..4004)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4104..4126
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4164..4182)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4250..4354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4355..4423,4424..5215)
                     /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     complement(4573..4592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5386..5974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5875..5894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
      721 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
      781 gaagttatgg cgcgccatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
      841 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
      901 attcggtcga acgcgtcgcc gcgtgtttaa acgcattagt cttccaattg aaaaaagtga
      961 tttaatttat accattttaa ttcagctttg taaaaatgta tcaaagagat agcaaggtat
     1021 tcagttttag taaacaagat aattgctcct aaagtagccc cttgaattcc gaggcagtag
     1081 gcagccgcga ttaggctgtt ataatacatc tgtggcttca ctattataac agcctaatcg
     1141 cggccgctca ctgtcaacag caatatacct tctcgagcct tctgttgggt taacctgaag
     1201 aagtaatccc agcaagtgtt tccaagatgt gcaggcaacg attctgtaaa gtactgaagc
     1261 ctcattcaac atagtatatg tgctgccgaa gcgagcactt aacaaggctt gcggccgcct
     1321 acaggaacag gtggtggcgg ccctcggcgc gctcgtactg ctccacgatg gtgtagtcct
     1381 cgttgtggga ggtgatgtcc agcttggagt ccacgtagta gtagccgggc agctgcacgg
     1441 gcttcttggc catgtagatg gacttgaact ccaccaggta gtggccgccg tccttcagct
     1501 tcagggcctt gtggatctcg cccttcagca cgccgtcgcg ggggtacagg cgctcggtgg
     1561 aggcctccca gcccatagtc ttcttctgca ttacggggcc gtcggagggg aagttcacgc
     1621 cgatgaactt caccttgtag atgaaggagc cgtcctgcag ggaggagtcc tgggtcacgg
     1681 tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
     1741 acagcttctt gtagtcgggg atgtcggcgg ggtgcttcac gtacaccttg gagccgtact
     1801 ggaactgggg ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca
     1861 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
     1921 cgtggccgtt cacggagccc tccatgcgca ccttgaagcg catgaactcc ttgatgacgt
     1981 cctcggagga gtccatcctg caggcccggt agcgctagca taacttcgta taaagtatcc
     2041 tatacgaagt tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa
     2101 agaataactt cgtataatgt atgctatacg aagttatgaa ttcgatatca agcttatcga
     2161 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2221 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2281 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2341 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2401 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2461 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2521 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2581 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2641 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2701 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     2761 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     2821 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     2881 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     2941 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     3001 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     3061 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     3121 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3181 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3241 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     3301 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3361 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3421 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3481 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3541 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3601 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3661 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3721 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3781 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3841 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3901 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3961 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4021 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4081 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4141 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4201 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4261 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4321 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4381 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4441 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4501 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4561 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4621 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4681 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4741 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4801 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4861 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4921 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4981 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5041 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5101 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5161 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5221 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5281 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5341 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5401 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5461 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5521 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5581 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5641 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5701 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5761 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5821 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5881 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5941 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6001 gttcctggcc ttttgctggc cttttgctca catgt
//
