LOCUS       sequence-362507-        6657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This vector simultaneously expresses mCherry protein and shRNA in a 
            Cre-dependent manner..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6657)
  AUTHORS   Kim SC, Kim A, Park JY, Hwang EM
  TITLE     Improved AAV vector system for cell-type-specific RNA interference.
  JOURNAL   J Neurosci Methods. 2021 Dec 21:109452. doi: 
            10.1016/j.jneumeth.2021.109452.
  PUBMED    34953938
REFERENCE   2  (bases 1 to 6657)
  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 Neurosci 
            Methods. 2021 Dec 21:109452. doi: 10.1016/j.jneumeth.2021.109452."
COMMENT     Sequence Label: pAAV-R-Creon shRNA[Control]
FEATURES             Location/Qualifiers
     source          1..6657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1408
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1399
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1356..1376
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1433..1466)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1543..1576)
                     /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."
     CDS             complement(1625..2335)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGTVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1903..1922)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2172..2190
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2332..2341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2358..2391
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2442..2475
                     /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     complement(2493..2513)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_feature    2784..3372
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2837..2857)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3255..3266)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3404..3880
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3603..3622)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3920..4060
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3920..4049
                     /label=AAV2 ITR
     rep_origin      4135..4590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4222..4241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4432..4453
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4607..4626)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4726..4748
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4786..4804)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4872..4976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4977..5045,5046..5837)
                     /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(5195..5214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6008..6596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6497..6516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccactg
      541 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
      601 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
      661 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
      721 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
      781 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
      841 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
      901 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
      961 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     1021 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     1081 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     1141 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     1201 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1261 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1321 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1381 tttttttctt ccatttcagg tgtcgtgagg taccggatcc tctagatccg gaataacttc
     1441 gtatagcata cattatacga agttatgcag aatggtagct ggattgtagt cgacgtgtgt
     1501 tttgagacta taaagctgct attagcaata tgaaacctct taataacttc gtatagtata
     1561 aattatacga agttataagc cttgttaacg cgcggtgacc gctagcacta gttactagga
     1621 tccgctactt gtacagctcg tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt
     1681 actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccaacttg atgttgacgt
     1741 tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcagcgt
     1801 cgtagtggcc gccgtccttc agcttcagcc tctgcttgat ctcgcccttc agggcgccgt
     1861 cctcggggta catccgctcg gaggaggcct cccagcccat cgtcttcttc tgcattacgg
     1921 ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct
     1981 gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct
     2041 cccacttgaa gccctcgggg aaggacagct tcaagtagtc ggggatgtcg gcggggtgct
     2101 tcacgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag gcgaagggca
     2161 gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     2221 cgccctcgcc ctcgatctcg aactcgtggc cgttcacggt gccctccatg tgcaccttga
     2281 agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc accatggtgg
     2341 cggtggcgac cggtagcata acttcgtata atgtatgcta tacgaagtta tttgccttaa
     2401 cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg tataatttat
     2461 actatacgaa gttataatta ctttacagtt agggtgagtt tccttttgtg ctgtttttta
     2521 aaataataat ttagtatttg tatctcttat agaaatccaa gcctatcatg taaaatgtag
     2581 ctagtattaa aaagaacaga ttatctgtct tttatcgcac attaagcctc tatagttact
     2641 aggaaatatt atatgcaaat taaccggggc aggggagtag ccgagcttct cccacaagtc
     2701 tgtgcgaggg ggccggcgcg ggcctagaga tggcggcgtc ggatctctag agcgggcccg
     2761 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2821 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2881 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2941 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3001 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3061 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3121 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3181 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     3241 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3301 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3361 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     3421 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     3481 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     3541 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     3601 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     3661 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     3721 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     3781 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     3841 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     3901 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3961 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4021 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     4081 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4141 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4201 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4261 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4321 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4381 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4441 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     4501 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4561 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4621 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4681 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4741 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4801 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4861 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4921 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4981 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5041 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5101 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5161 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5221 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5281 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5341 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5401 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5461 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5521 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5581 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5641 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5701 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5761 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5821 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5881 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5941 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
