LOCUS       Exported                6443 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Per2 transcription fluorescence reporter.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-P(Per2)-DIO-mCherry-Venus-NLS-D2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  AUTHORS   Mei L, Fan Y, Lv X, Welsh DK, Zhan C, Zhang EE
  TITLE     Long-term in vivo recording of circadian rhythms in brains of freely
            moving mice.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Apr 2. pii: 1717735115. doi: 
            10.1073/pnas.1717735115.
  PUBMED    29610316
REFERENCE   2  (bases 1 to 6443)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6443
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   43..172
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    731..764
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    815..848
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      857..866
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             863..1573
                     /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(1008..1026)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1276..1295
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(1722..1742)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1746..2462)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1788..1809)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2137..2156
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2396..2417
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2459..2468
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(2470..2503)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(2554..2587)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2612..3200
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2665..2685)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3083..3094)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3232..3708
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3431..3450)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3748..3888
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3748..3877
                     /label=AAV2 ITR
     rep_origin      3963..4418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4050..4069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4260..4281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4435..4454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4554..4576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4614..4632)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4700..4804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4805..5665
                     /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(5023..5042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5836..6424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6325..6344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
       61 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      121 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
      181 acgcgtcgga ttaccgaggc tggtcacgtc gtcgcaggtg ataggccggg ggccctggtc
      241 tctgccggct gtgagttgcg cagcggccaa gcaccattcc cccgcgccgc agtggtacgc
      301 gccactccgg ggctgcacga gcgggccacc gccgtgccag gtgaatggaa gtcccgcagg
      361 ccggaagtgg acgagcctac tcgcccgggc gcgggggggc gcaagagcgc gcagcatctt
      421 cattgaggaa cccgggcggc gaacatggag ttccatgtgc gtcttatgta aagagagcga
      481 cgggcgtctc caccaattga cgagcgtagc tctcaggttc cgccccgcca gtatgcaaat
      541 gaggtggcac tccgaccaat ggcgcgcgca ggggcgggct cagcgcgcgc ggtcacgttt
      601 tccactatgt gacagcggag ggcgacgcgg cggcagcggc gctactggga ctagcggctc
      661 cgggcggctg cggcgcaggc cgagcgcacc aagtgacggg ccgagcaagg gatctagagt
      721 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
      781 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
      841 gaagttatgg cgcgccgcca ccatggtgag caagggcgag gaggataaca tggccatcat
      901 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      961 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     1021 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     1081 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     1141 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     1201 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     1261 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     1321 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     1381 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     1441 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     1501 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     1561 gctgtacaag taggctacac attgatccta gcagaagcac aggctgcagg gtgacggtcc
     1621 atcccgctct cctgggcaca agacatgggc agcgtgccat catcctgctc ctccacctcc
     1681 ggcgggaagc catggctaag cttagaattc acggggtctt ctacctttct cttctttttt
     1741 ggaggcttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     1801 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gtttggactg gtagctcagg
     1861 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     1921 tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt tgccttgatg
     1981 ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc
     2041 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     2101 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     2161 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     2221 tgcttcatgt ggtcggggta gcgggcgaag cactgcaggc cgtagccgag ggtggtcacg
     2281 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tcaacttcag ggtcagcttg
     2341 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     2401 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     2461 atggtggcca taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa
     2521 ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg
     2581 aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     2641 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2701 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2761 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2821 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2881 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2941 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3001 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     3061 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3121 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3181 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     3241 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3301 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3361 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3421 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3481 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3541 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3601 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3661 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     3721 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3781 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3841 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3901 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3961 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4021 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4081 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4141 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4201 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4261 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4321 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4381 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4441 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4501 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4561 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4621 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4681 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4741 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4801 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4861 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4921 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4981 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5041 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5101 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5161 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5221 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5281 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5341 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5401 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5461 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5521 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5581 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5641 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5701 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5761 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5821 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5881 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5941 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6001 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6061 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6121 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6181 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6241 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6301 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6361 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6421 gaaaaacgcc agcaacgcgg cct
//
