LOCUS       sequence-171362-        7461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-inducible shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7461)
  AUTHORS   Jung JY, Lee SE, Hwang EM, Lee CJ
  TITLE     Neuronal Expression and Cell-Type-Specific Gene-Silencing of Best1 
            in Thalamic Reticular Nucleus Neurons Using pSico-Red System.
  JOURNAL   Exp Neurobiol. 2016 Jun;25(3):120-9. doi: 10.5607/en.2016.25.3.120. 
            Epub 2016 Jun 22.
  PUBMED    27358580
REFERENCE   2  (bases 1 to 7461)
  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: 
            "10.5607/en.2016.25.3.120"; journalName: "Exp Neurobiol"; date: 
            "2016-06"; volume: "25"; issue: "3"; pages: "120-9"
COMMENT     Sequence Label: pAAV-Sico-Red
FEATURES             Location/Qualifiers
     source          1..7461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   71..211
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   71..200
                     /label=AAV2 ITR
     rep_origin      286..741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(373..392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(758..777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     877..899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(937..955)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1023..1127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1128..1196,1197..1988)
                     /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(1346..1365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2159..2747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2648..2667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2809..2938
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2953..4131
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3184..4122
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4079..4099
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4157..4867
                     /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(4302..4320)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4570..4589
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4989..5110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5026..5045)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5080..5099
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5378..5398
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(5416..5449)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     enhancer        5549..5852
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5853..6056
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6007..6027
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6059..6079
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      6087..6096
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6093..6095,6096..6098,6099..6812)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6093..6095,6096..6098,6099..6809)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6138..6159)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6399..6418)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6746..6767
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6820..6853)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     primer_bind     6921..6937
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6922..6938
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6983..7007)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    join(7016..7461,1..31)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(7215..7234)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
       61 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
      121 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
      181 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
      241 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
      301 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      361 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      421 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      481 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
      541 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      601 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
      661 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      721 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
      781 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      841 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      901 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      961 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1141 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1201 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1261 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1321 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1381 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1441 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1501 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1561 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1621 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1681 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1741 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1801 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1861 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1921 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1981 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     2821 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     2881 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     2941 ggccgcacgc gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     3001 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     3061 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     3121 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     3181 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
     3241 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
     3301 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
     3361 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
     3421 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     3481 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     3541 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     3601 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     3661 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     3721 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     3781 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     3841 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     3901 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     3961 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     4021 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     4081 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg attcgaacat
     4141 cgattgaatt cccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     4201 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     4261 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     4321 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     4381 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     4441 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     4501 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     4561 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     4621 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     4681 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     4741 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     4801 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     4861 caagtaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     4921 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4981 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5041 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5101 atgtatctta aggcgtacaa gttgtacaaa aaagcaggct tctagagatc cgacgcgcca
     5161 tctctaggcc cgcgccggcc ccctcgcaca gacttgtggg agaagctcgg ctactcccct
     5221 gccccggtta atttgcatat aatatttcct agtaactata gaggcttaat gtgcgataaa
     5281 agacagataa tctgttcttt ttaatactag ctacatttta catgataggc ttggatttct
     5341 ataagagata caaatactaa attattattt taaaaaacag cacaaaagga aactcaccct
     5401 aactgtaaag taattataac ttcgtatagt ataaattata cgaagttata agccttggtt
     5461 ttttgaattc cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     5521 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     5581 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     5641 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     5701 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     5761 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     5821 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     5881 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     5941 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     6001 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggca
     6061 actttgtata caaaagttga ccggtcgcca ccatggtgag caagggcgag gagctgttca
     6121 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     6181 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     6241 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     6301 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     6361 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     6421 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     6481 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     6541 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     6601 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     6661 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     6721 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     6781 tcactctcgg catggacgag ctgtacaagt agcggccgca taacttcgta tagtataaat
     6841 tatacgaagt tataagcctt gttttttgcc aacttgtcaa tgaattcaag agattcattg
     6901 acaagttggc aaattttttc tcgaggtcga cggtatcgat aagctcgctt cacgagatca
     6961 tgtttaaggg ttcccgttcc aaacccagct ttcttgtaca aagtggtaga tctacgggtg
     7021 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     7081 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     7141 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     7201 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     7261 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     7321 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     7381 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     7441 tcccaaattg ctgggattac a
//
