LOCUS       Exported                6800 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV transfer vector with mammalian codon-optimized ChIEF-tdTomato 
            between AAV2 ITR.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-oChIEF-tdTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6800)
  AUTHORS   Lin JY, Lin MZ, Steinbach P, Tsien RY
  TITLE     Characterization of engineered channelrhodopsin variants with 
            improved properties and kinetics.
  JOURNAL   Biophys J. 2009 Mar 4;96(5):1803-14. doi: 10.1016/j.bpj.2008.11.034.
  PUBMED    19254539
REFERENCE   2  (bases 1 to 6800)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6800
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1563..2993
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    3027..3615
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3080..3100)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3498..3509)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3618..3634)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3619..3635)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3664..3785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3675..3694)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3729..3748
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3947..4076
                     /label=AAV2 ITR
     primer_bind     complement(4142..4159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4313..4901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4393..4412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5072..5932)
                     /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     5695..5714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5933..6037)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6063..6518)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6200..6221)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6412..6431
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6590..6719
                     /label=AAV2 ITR
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gaattcaagc tgctagcaag gatccgacca
      481 tggtgagcag aagaccctgg ctgctggccc tggccctggc cgtggccctg gccgccggca
      541 gcgccggcgc cagcaccggc agcgacgcca ccgtgcccgt ggccacccag gacggccccg
      601 actacgtgtt ccacagagcc cacgagagaa tgctgttcca gaccagctac accctggaga
      661 acaacggcag cgtgatctgc atccccaaca acggccagtg cttctgcctg gcctggctga
      721 agagtaacgg caccaacgcc gagaagctgg ccgccaacat cctgcagtgg atcaccttcg
      781 ccctgagcgc cctgtgcctg atgttctacg gctaccagac ctggaagagt acctgcggct
      841 gggaggagat ctacgtggcc accatcgaga tgatcaagtt catcatagag tacttccacg
      901 agttcgacga gcccgccgtg atctacagca gcaacggcaa caagaccgtg tggctgagat
      961 acgccgagtg gctgctgacc tgccccgtgg tcctgatcca cctgagcaac ctgaccggcc
     1021 tggccaacga ctacaacaag agaaccatgg gcctgctggt gagcgacatc ggcaccatcg
     1081 tgtggggcac caccgccgcc ctgagcaagg gctacgtgag agtgatcttc ttcctgatgg
     1141 gcctgtgcta cggcatctac accttcttca acgccgccaa ggtgtacatc gaggcctacc
     1201 acaccgtgcc caagggcaga tgcagacagg tggtgaccgg catggcctgg ctgttcttcg
     1261 tgagctgggg catgttcccc atcctgttca tcctgggccc cgagggcttc ggcgtgctga
     1321 gcgtgtacgg cagcaccgtg ggccacacca tcatcgacct gatgagcaag aactgctggg
     1381 gcctgctggg ccactacctg agagtgctga tccacgagca catcctgatc cacggcgaca
     1441 tcagaaagac caccaagctg aacatcggcg gcaccgagat cgaggtggag accctggtgg
     1501 aggacgaggc cgaggccggc gccgtgaaca agggcaccgg caagtacgag agcagcatcg
     1561 atatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg
     1621 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1681 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     1741 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca
     1801 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1861 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     1921 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     1981 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     2041 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     2101 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc
     2161 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     2221 gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc agctccggca
     2281 ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc ttcaaggtgc
     2341 gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2401 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
     2461 cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg
     2521 ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2581 tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca
     2641 cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc
     2701 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     2761 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     2821 tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca
     2881 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct
     2941 ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag taggctagcc
     3001 ccgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3061 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3121 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3181 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3241 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3301 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3361 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3421 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3481 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3541 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3601 gggccgcctc cccgcatcga taccgtcgac ctcgacccgg gcggccgctt cgagcagaca
     3661 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     3721 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3781 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     3841 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag
     3901 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     3961 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4021 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4081 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4141 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4201 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4261 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4321 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4381 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4441 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4501 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4561 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4621 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4681 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4741 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4801 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4861 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4921 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4981 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5041 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5101 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5161 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5221 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5281 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5341 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5401 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5461 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5521 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5581 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5641 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5701 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5761 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5821 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5881 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5941 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6001 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6061 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     6121 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     6181 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     6241 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     6301 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     6361 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     6421 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     6481 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     6541 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg
     6601 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
     6661 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
     6721 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagatctc
     6781 tgcagagggc cctgcgtatg
//
