LOCUS       Exported                5852 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Slow cycling (open for minutes) step-function artificial anion 
            conducting channelrhodopsin (aACR). High light sensitivity. 
            Activation with green light, inactivation max with 625 nm 
            (accelarates closure to ms). Not recommended for in vivo use. Codon 
            optimized for mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    AAV2_hSyn_Aurora_CA_Citrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5852)
  AUTHORS   Wietek J, Rodriguez-Rozada S, Tutas J, Tenedini F, Grimm C, Oertner 
            TG, Soba P, Hegemann P, Wiegert JS
  TITLE     Anion-conducting channelrhodopsins with tuned spectra and modified 
            kinetics engineered for optogenetic manipulation of behavior.
  JOURNAL   Sci Rep. 2017 Nov 2;7(1):14957. doi: 10.1038/s41598-017-14330-y.
  PUBMED    29097684
REFERENCE   2  (bases 1 to 5852)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5852
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   29..158
                     /label=AAV2 ITR
     promoter        258..705
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1649..2365
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1694..1715)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1955..1974)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2302..2323
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2381..2969
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2434..2454)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2852..2863)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3010..3217
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3433..3450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3604..4192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3684..3703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4363..5223)
                     /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     4986..5005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5224..5328)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5354..5809)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5491..5512)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5703..5722
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg
       61 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
      121 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc
      181 atgctactta tctacgtagc catgctctag gaagatcgta ggttaattaa tctagactgc
      241 agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt
      301 gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt
      361 gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg
      421 ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc
      481 ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc
      541 caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag
      601 ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag
      661 tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag aggatccacc
      721 ggtgaccatg gattacggcg gagccctgtc agcagtcggc ctgtttcaga cctcatatac
      781 cctggagaat aacggcagcg tcatctgtat cccaaataac gggcagtgtt tctgcctggc
      841 atggctgaag agcaacggca ccaatgcaga gaagctggca gccaatatcc tgcagtgggt
      901 gtcatttgcc ctgtctgtgg catgtctggg atggtacgcc tatcaggcat ggagagccac
      961 ttgtggatgg gagaacgtgt atgtcgctct gattcagatg atgaagagca tcattgaggc
     1021 cttccattcc tttgattcac cagctactct gtggctgtca agtggcaacg gggtcaggtg
     1081 gatgagatat gggagctggc tgctgaccgc ccccgtgatt ctgatccacc tgagcaacct
     1141 gactggcctg aaggacgatt acagcaagcg cactatgggg ctgctggtct cagacgtggg
     1201 atgtatcgtg tggggcgcca caagcgcaat gtgcacaggc tggaccaaga tcctgttttt
     1261 cctgatcagc ctgtcttacg gcatgtacac ttacttccat gccgcaaagg tgtacatcga
     1321 ggctttccac accgtgccaa agggactgtg cagacagctg gtgcgggcaa tggcttggct
     1381 gttctttgtg agctggggga tgtttccagt cctgtttctg ctgggacccg agggcttcgg
     1441 gcacatcagc agatacgggt ccaacattgg ccacagcatt ctggatctga ttgccaagca
     1501 gatgtgggga gtgctgggaa actacctgag ggtgaaaatt cacagccata tcctgctgta
     1561 cggagatatc cgcaagaagc agaaaatcac cattgcaggg caggaaatgg aagtggaaac
     1621 tctggtcgcc gaggaggaag acctcgagat ggtgagcaag ggcgaggagc tgttcaccgg
     1681 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1741 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1801 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg
     1861 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1921 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1981 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2041 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2101 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2161 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2221 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     2281 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2341 tctcggcatg gacgagctgt acaagtagaa gcttatcgat aatcaacctc tggattacaa
     2401 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2461 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2521 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2581 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2641 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2701 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2761 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     2821 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2881 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2941 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgctcat
     3001 aaagctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3061 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3121 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3181 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtccggact gtacaattgc
     3241 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3301 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     3361 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     3421 tctgcgcgct cgctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5341 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5401 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5461 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5521 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5581 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5641 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5701 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5761 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     5821 ttcaggctgc gcaactgttg ggaagggcga tc
//
