LOCUS       Exported                6001 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Red-shifted artificial anion conducting channelrhodopsin (aACR). 
            Activation up to 600 nm; off-kinetics 260 ms. Codon optimized for 
            mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    AAV2_hSyn_Aurora_Citrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  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 6001)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6001
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   13..142
                     /label=AAV2 ITR
     promoter        242..689
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1633..2349
                     /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(1678..1699)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1939..1958)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2286..2307
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2365..2953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2418..2438)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2836..2847)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2994..3201
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3371..3500
                     /label=AAV2 ITR
     primer_bind     complement(3566..3583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3737..4325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3817..3836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4496..5356)
                     /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     5119..5138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5357..5461)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5487..5942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5624..5645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5836..5855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg
      181 tagccatgct ctaggaagat cgtaggttaa ttaatctaga ctgcagaggg ccctgcgtat
      241 gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg
      301 accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag
      361 agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc
      421 ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg
      481 cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc
      541 gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc
      601 gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg
      661 gaggagtcgt gtcgtgcctg agagcgcagt cgagaggatc caccggtgac catggattac
      721 ggcggagccc tgtcagcagt cggcctgttt cagacctcat ataccctgga gaataacggc
      781 agcgtcatct gtatcccaaa taacgggcag tgtttctgcc tggcatggct gaagagcaac
      841 ggcaccaatg cagagaagct ggcagccaat atcctgcagt gggtgtcatt tgccctgtct
      901 gtggcatgtc tgggatggta cgcctatcag gcatggagag ccacttgtgg atgggagaac
      961 gtgtatgtcg ctctgattca gatgatgaag agcatcattg aggccttcca ttcctttgat
     1021 tcaccagcta ctctgtggct gtcaagtggc aacggggtca ggtggatgag atatgggagc
     1081 tggctgctga cctgccccgt gattctgatc cacctgagca acctgactgg cctgaaggac
     1141 gattacagca agcgcactat ggggctgctg gtctcagacg tgggatgtat cgtgtggggc
     1201 gccacaagcg caatgtgcac aggctggacc aagatcctgt ttttcctgat cagcctgtct
     1261 tacggcatgt acacttactt ccatgccgca aaggtgtaca tcgaggcttt ccacaccgtg
     1321 ccaaagggac tgtgcagaca gctggtgcgg gcaatggctt ggctgttctt tgtgagctgg
     1381 gggatgtttc cagtcctgtt tctgctggga cccgagggct tcgggcacat cagcagatac
     1441 gggtccaaca ttggccacag cattctggat ctgattgcca agcagatgtg gggagtgctg
     1501 ggaaactacc tgagggtgaa aattcacagc catatcctgc tgtacggaga tatccgcaag
     1561 aagcagaaaa tcaccattgc agggcaggaa atggaagtgg aaactctggt cgccgaggag
     1621 gaagacctcg agatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1681 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1741 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1801 ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc
     1861 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1921 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1981 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2041 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2101 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2161 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2221 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     2281 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2341 ctgtacaagt agaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2401 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2461 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2521 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2581 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2641 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2701 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2761 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2821 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2881 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2941 gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc tcataaagct cgactgtgcc
     3001 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3061 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3121 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3181 caatagcagg catgctgggg atgcggtccg gactgtacaa ttgcattcat tttatgtttc
     3241 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3301 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     3361 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3421 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3481 gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3541 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3601 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3661 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4141 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4321 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4441 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4501 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4561 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4621 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4681 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4741 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4801 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4861 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4921 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4981 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5041 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5101 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5161 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5221 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5281 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5341 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5401 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5461 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     5521 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5581 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5641 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5701 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5761 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5821 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5881 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5941 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6001 g
//
