LOCUS       Exported                7089 bp ds-DNA     circular SYN 13-MAY-2022
DEFINITION  Optogenetics.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CaMKIIa-ChRmine-oScarlet-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7089)
  AUTHORS   Kishi KE, Kim YS, Fukuda M, Inoue M, Kusakizako T, Wang PY, 
            Ramakrishnan C, Byrne EFX, Thadhani E, Paggi JM, Matsui TE, 
            Yamashita K, Nagata T, Konno M, Quirin S, Lo M, Benster T, Uemura T,
            Liu K, Shibata M, Nomura N, Iwata S, Nureki O, Dror RO, Inoue K, 
            Deisseroth K, Kato HE
  TITLE     Structural basis for channel conduction in the pump-like 
            channelrhodopsin ChRmine.
  JOURNAL   Cell. 2022 Feb 17;185(4):672-689.e23. doi: 
            10.1016/j.cell.2022.01.007. Epub 2022 Feb 2.
  PUBMED    35114111
REFERENCE   2  (bases 1 to 7089)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7089
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1470..1479
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2472..3164
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWDRVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2602..2620)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2870..2889
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3216..3804
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3269..3289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3687..3698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3836..4312
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4035..4054)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4352..4492
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4352..4481
                     /label=AAV2 ITR
     rep_origin      4567..5022
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4654..4673)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4864..4885
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5039..5058)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5158..5180
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5218..5236)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5304..5408
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5409..6269
                     /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(5627..5646)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6440..7028
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6929..6948
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccccg ggtaccggtg ccaccatggc acacgcacca ggcaccgacc
     1501 agatgttcta cgtgggcaca atggacggct ggtatctgga taccaagctg aactccgtgg
     1561 ccatcggcgc ccactggtct tgctttatcg tgctgacaat caccacattc tacctgggct
     1621 atgagtcctg gacatctcgg ggccctagca agagaacctc cttttacgcc ggctatcagg
     1681 aggagcagaa cctggccctg tttgtgaatt tctttgccat gctgagctac ttcggcaaga
     1741 tcgtggccga cacactgggc cacaacttcg gcgatgtggg ccccttcatc atcggcttcg
     1801 gcaattacag gtatgccgac tacatgctga cctgcccaat gctggtgtac gatctgctgt
     1861 atcagctgag ggccccctat cgcgtgtctt gtagcgccat catctttgcc atcctgatgt
     1921 ctggcgtgct ggccgagttc tacgcagagg gcgaccctag gctgaggaat ggcgcctacg
     1981 cctggtatgg ctttggctgt ttctggttta tcttcgccta ctctatcgtg atgagcatcg
     2041 tggccaagca gtatagccgg ctggcccagc tggcccagga tacaggcgcc gagcacagcc
     2101 tgcacgtgct gaagttcgcc gtgtttacct tctccatgct gtggattctg tttcccctgg
     2161 tgtgggccat ctgccctaga ggcttcggct ggatcgacga taactggaca gaggtggccc
     2221 actgcgtgtg cgacatcgtg gccaagtcct gttacggctt tgccctggcc cggttcagaa
     2281 agacctatga tgaggagctg tttcggctgc tggagcagct gggacacgac gaggatgagt
     2341 tccagaagct ggagctggat atgaggctga gcagcaatgg cgagcgcctg cggagactgt
     2401 ctgcggccgc caagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg
     2461 acatcaacgt ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     2521 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2581 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     2641 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
     2701 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gaccgcgtga
     2761 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     2821 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     2881 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     2941 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cgggcgctac ctggcggact
     3001 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtggacc
     3061 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     3121 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagttctgc tacgagaacg
     3181 aggtgtaatg agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     3241 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3301 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3361 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3421 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3481 agctcctttc cggaactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3541 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3601 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     3661 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3721 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3781 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     3841 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     3901 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     3961 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     4021 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     4081 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     4141 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     4201 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     4261 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     4321 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4381 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4441 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4501 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4561 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4621 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     4681 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4741 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4801 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4861 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     4921 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4981 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     5041 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5101 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5161 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5221 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5281 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5341 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5401 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5461 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5521 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5581 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5641 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5701 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5761 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5821 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5881 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5941 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6001 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6061 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6121 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6181 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6241 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6301 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6361 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6421 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6481 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6541 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6601 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6661 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6721 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6781 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6841 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6901 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6961 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7021 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7081 ctcacatgt
//
