LOCUS       Exported                5598 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses an optimized mosquito OPN3 rhodopsin in-frame with 
            mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5598)
  AUTHORS   Mahn M, Saraf-Sinik I, Patil P, Pulin M, Bitton E, Karalis N, 
            Bruentgens F, Palgi S, Gat A, Dine J, Wietek J, Davidi I, Levy R, 
            Litvin A, Zhou F, Sauter K, Soba P, Schmitz D, Luthi A, Rost BR, 
            Wiegert JS, Yizhar O
  TITLE     Efficient optogenetic silencing of neurotransmitter release with a 
            mosquito rhodopsin
  JOURNAL   Neuron
REFERENCE   2  (bases 1 to 5598)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5598
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   24..153
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1661..2356
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1794..1812)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2062..2081
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2396..2799
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    2806..2854
                     /note="synthetic polyadenylation signal"
     repeat_region   2884..3024
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2884..3013
                     /label=AAV2 ITR
     rep_origin      3099..3554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3186..3205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3396..3417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3571..3590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3690..3712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3750..3768)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3836..3940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3941..4801
                     /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(4159..4178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4972..5560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5461..5480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
      121 agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtctt gtggactaag
      181 tttgttcgca tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac
      241 ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag
      301 caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
      361 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
      421 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt
      481 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc tctagagcgg
      541 taccggatcc gccaccatgt acgacgcccc caacgacgtg gccagcagcg tggccgacta
      601 cgaggacctg atggccccct gggcctacaa cgccgccgcc atcaccctgt tcttcatcgg
      661 cttcttcggc ttcttcctga acctgttcgt gatcgccctg atgagcaagg acatgcagct
      721 gtggaccccc atgaacatca tcctgttcaa cctggtgtgc agcgacttca gcgtgagcat
      781 catcggcaac cccctgaccc tgaccagcgc catcagccac aggtggatct tcggcaggac
      841 cctgtgcgtg gcctacggct tcttcatgag cctgctgggc atcaccagca tcaccaccct
      901 gaccgtgctg agctacgaga ggtactgcct gatcagcagg cccttcagca gcaggaacct
      961 gagcaggaag ggcgccttcc tggccatctt cttcatctgg ggctacagct tcgccctgac
     1021 cagccccccc ctgttcggct ggggcgccta cgtgcaggag gccgccaaca tcagctgcag
     1081 cgtgaactgg gagagccaga ccaagaacgc caccacctac atcatcttcc tgttcgtgtt
     1141 cggcctggtg gtgcccctga tcgtgatcgt gtacagctac accaacatca tcgtgtacat
     1201 gaggaggaac agcgccaggg tgggcaggat caacagggcc gagcagaggg tgaccagcat
     1261 ggtggccgtg atgatcgtgg ccttcatggt ggcctggacc ccctacgcca tcttcgccct
     1321 gatcgagcag ttcggccccc ccgagctgat cggccctgga ctggctgtgc tgcctgctct
     1381 gatcgccaag agcagcatct gctacaaccc catcatctac gtgggcatga acacccagtt
     1441 cagggccgcc ttcaccaggg tgaggaacaa gggcggcgtg cccaccgccg accagaacac
     1501 caccaccatg cagagggagc tgaccaagag cagcagggac atggtgaccg agaccagcca
     1561 ggtggctcct gctccgcgag cccgagatcc aaccggtaag agcaggatca ccagcgaggg
     1621 cgagtacatc cccctggacc agatcgacat caacgtggtg atggtgagca agggcgaggc
     1681 agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga
     1741 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     1801 gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat
     1861 gtacggctcc agggccttca ccaagcaccc cgccgacatc cccgactact ataagcagtc
     1921 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac
     1981 cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg
     2041 caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc
     2101 caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg
     2161 cctgaaggac ggcggacgct acctggcgga cttcaagacc acctacaagg ccaagaagcc
     2221 cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga
     2281 ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca ccggcggcat
     2341 ggacgagctg tacaagttct gctacgagaa cgaggtgtaa ctagaattca agcttaatca
     2401 acctctggat tacaaaattt gtgaaagatt gactgatatt cttaactatg ttgctccttt
     2461 tacgctgtgt ggatatgctg ctttaatgcc tctgtatcat gctattgctt cccgtatggc
     2521 tttcgttttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2581 cgttgtccgt caacgtggcg tggtgtgctc tgtgtttgct gacgcaaccc ccactggctg
     2641 gggcattgcc accacctgtc aactcctttc tgggactttc gctttccccc tcccgatcgc
     2701 cacggcagaa ctcatcgccg cctgccttgc ccgctgctgg acaggggcta ggttgctggg
     2761 cactgataat tccgtggtgt tgtcggggaa gctgacgtca gcgctaataa aatcgctatc
     2821 catcgaagat ggatgtgtgt tggttttttg tgtgggtaac ccacgtgcgg accgagcggc
     2881 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2941 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3001 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     3061 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3121 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     3181 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3241 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3301 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3361 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3421 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3481 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3541 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3601 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3661 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3721 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3781 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3841 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3901 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3961 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4021 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4081 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4141 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4201 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4261 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4321 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4381 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4441 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4501 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4561 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4621 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4681 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4741 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4801 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4861 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4921 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4981 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5041 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5101 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5161 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5221 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5281 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5341 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5401 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5461 gggaaacgcc tggtatcttt atagtcctgt cgggttttgc cacctctgac ttgagcgtcg
     5521 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5581 tttacggttc ctggcctt
//
