LOCUS       Exported                7065 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encoding aCamkII-mCherry-Cre.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-aCamkII-mCherry-P2A-Cre-WPRE-BGH-polyA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Zhou H, Liu J, Zhou C, Gao N, Rao Z, Li H, Hu X, Li C, Yao X, Shen 
            X, Sun Y, Wei Y, Liu F, Ying W, Zhang J, Tang C, Zhang X, Xu H, Shi 
            L, Cheng L, Huang P, Yang H
  TITLE     In vivo simultaneous transcriptional activation of multiple genes in
            the brain using CRISPR-dCas9-activator transgenic mice.
  JOURNAL   Nat Neurosci. 2018 Jan 15. pii: 10.1038/s41593-017-0060-6. doi: 
            10.1038/s41593-017-0060-6.
  PUBMED    29335603
REFERENCE   2  (bases 1 to 7065)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7065
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        812..2100
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      2126..2135
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2132..2839
                     /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(2277..2295)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2545..2564
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2855..2911
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2912..2932
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3959..3985
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    4019..4607
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4072..4092)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4490..4501)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4643..4660)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4649..4873
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4978..5118
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4978..5107
                     /label=AAV2 ITR
     rep_origin      5193..5648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5280..5299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5490..5511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5665..5684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5784..5806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5844..5862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5930..6034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6035..6895
                     /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(6253..6272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcaa tcgatttcat atgccggtta acattatggc cttaggtcac ttcatctcca
      841 tggggttctt cttctgattt tctagaaaat gagatggggg tgcagagagc ttcctcagtg
      901 acctgcccag ggtcacatca gaaatgtcag agctagaact tgaactcaga ttactaatct
      961 taaattccat gccttggggg catgcaagta cgatatacag aaggagtgaa ctcattaggg
     1021 cagatgacca atgagtttag gaaagaagag tccagggcag ggtacatcta caccacccgc
     1081 ccagccctgg gtgagtccag ccacgttcac ctcattatag ttgcctctct ccagtcctac
     1141 cttgacggga agcacaagca gaaactggga caggagcccc aggagaccaa atcttcatgg
     1201 tccctctggg aggatgggtg gggagagctg tggcagaggc ctcaggaggg gccctgctgc
     1261 tcagtggtga cagatagggg tgagaaagca gacagagtca ttccgtcagc attctgggtc
     1321 tgtttggtac ttcttctcac gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca
     1381 gggagagctg gaaagaaaca aggacagaga cagaggccaa gtcaaccaga ccaattccca
     1441 gaggaagcaa agaaaccatt acagagacta caagggggaa gggaaggaga gatgaattag
     1501 cttcccctgt aaaccttaga acccagctgt tgccagggca acggggcaat acctgtctct
     1561 tcagaggaga tgaagttgcc agggtaacta catcctgtct ttctcaagga ccatcccaga
     1621 atgtggcacc cactagccgt taccatagca actgcctctt tgccccactt aatcccatcc
     1681 cgtctgttaa aagggcccta tagttggagg tgggggaggt aggaagagcg atgatcactt
     1741 gtggactaag tttgttcgca tccccttctc caaccccctc agtacatcac cctgggggaa
     1801 cagggtccac ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg
     1861 caaagaggag caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg
     1921 caacgggaac agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct
     1981 cgcgaggctg tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc
     2041 ggttctccgt ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc
     2101 gctcgtaatc acctccgctg ctcgcgccac catggtgagc aagggcgagg aggataacat
     2161 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     2221 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     2281 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     2341 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2401 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2461 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     2521 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2581 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     2641 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     2701 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2761 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     2821 catggacgag ctgtacaagg ctagcggaag cggagccact aacttctccc tgttgaaaca
     2881 agcaggggat gtcgaagaga atcccgggcc acccaagaag aagaggaagg tgtccaatct
     2941 cctgactgtt caccagaacc tccctgcgct gccagtagat gccactagcg atgaggtcag
     3001 gaaaaatctc atggatatgt ttagggatag acaggcgttt tctgaacaca cctggaaaat
     3061 gctgcttagc gtgtgccgat cctgggcagc ctggtgtaag ctgaacaatc gcaaatggtt
     3121 ccccgccgag ccggaggacg tgcgcgatta cctgctgtat ctccaggcaa gagggctggc
     3181 tgtcaagact atccagcagc acttgggcca actgaatatg ctgcatcgac gcagcgggct
     3241 cccccggcct agcgattcaa acgcagtctc ccttgttatg aggagaatta gaaaggaaaa
     3301 cgtagatgcg ggtgagaggg ctaagcaggc tctcgctttt gagcggactg atttcgacca
     3361 ggtcagatcc ctgatggaga acagcgatcg gtgccaggac atcaggaacc tcgcatttct
     3421 gggaattgca tataacacac ttctgcgcat agctgagatc gcccggatca gagtgaaaga
     3481 catcagtcga acggacggcg gccggatgct tattcatatt ggacgcacaa agacattggt
     3541 cagcaccgct ggcgttgaaa aggccttgtc cctgggcgta acgaagctgg tggaaagatg
     3601 gatctcagtg tccggcgtgg ctgacgaccc taataattac ttgttctgtc gagtgagaaa
     3661 aaacggagtc gccgcgccct ctgccaccag ccaattgagt acacgggccc ttgaagggat
     3721 ctttgaggca acccaccgac tcatatacgg agccaaggat gacagtggcc agaggtatct
     3781 cgcctggtca ggtcattctg ctagggtggg ggccgcacga gacatggcgc gggcaggagt
     3841 ctccatacca gagattatgc aagctggagg ttggacaaat gtgaacatcg ttatgaacta
     3901 tatccgcaat cttgactctg aaaccggggc catggtgaga ctgctcgaag atggtgacta
     3961 cccatacgat gttccagatt acgcttaaga attcgatatc aagcttatcg ataagtaaaa
     4021 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4081 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4141 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4201 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4261 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4321 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4381 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     4441 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4501 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtctccg
     4561 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgccac cggttagggg
     4621 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4681 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4741 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4801 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4861 gtgggctcta tggcggcgcg tccagtgtgc tggaaatacc gagcgctaat aaaagatctt
     4921 tattttcatt agatctgtgt gttggttttt tgtgtcacgt gcggaccgag cggccgcagg
     4981 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     5041 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     5101 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5161 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     5221 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     5281 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5341 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5401 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5461 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5521 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     5581 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     5641 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     5701 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5761 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5821 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5881 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5941 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6001 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6061 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6121 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6181 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6241 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6301 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6361 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6421 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6481 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6541 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6601 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6661 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6721 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6781 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6841 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6901 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6961 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
