LOCUS       Exported                5852 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Blue-shifted artificial anion conducting channelrhodopsin (aACR). 
            Activation max 466 nm; off-kinetics 10 ms. Codon optimized for 
            mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    AAV2_hSyn_Phobos_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 12, 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   1..130
                     /label=AAV2 ITR
     promoter        230..677
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      697..706
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1642..2358
                     /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(1687..1708)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1948..1967)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2295..2316
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2374..2962
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2427..2447)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2845..2856)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3003..3210
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3576..4164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3656..3675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4335..5195)
                     /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     4958..4977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5196..5300)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5326..5781)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5463..5484)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5675..5694
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg taggttaatt aatctagact gcagagggcc ctgcgtatga gtgcaagtgg
      241 gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac
      301 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      361 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      421 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      481 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      541 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      601 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
      661 cgtgcctgag agcgcagtcg aattcaagct gctagcgcca ccatggacta tggcggcgct
      721 ttgtctgccg tcggactctt ccaaacctca tacactcttg agaacaatgg ttctgttatt
      781 tgcatcccga acaacggcca gtgcttctgc ttggcttggc ttaaatccaa cggaacaaat
      841 gccgagaagt tggctgccaa cattctgcag tggatttctt ttgcgctttc agcgctctgc
      901 ctgatgttct acggctacca gacctggaag tctacttgcg gctgggagaa catttacgtg
      961 gccacgatcc agatgatcaa gttcatcatc gagtatttcc atagctttga cgaacctgcg
     1021 gtgatctact catccaacgg caacaagacc cggtggcttc gttacgcgag ctggctgctg
     1081 acctgccctg tcattcttat ccacctgagc aaccttacgg gtctggcgaa cgactataac
     1141 aagcgtacca tgggtctgct ggtgtcagat atcggcggca tcgtgtgggc caccacggcc
     1201 gcgctgtcca agggatacgt ccgtgtcatt ttcttcctga tgggcctgtg ctacggcatc
     1261 tacacattct tcaacgcagc caaggtctac attgaggcgt accacaccgt gccgaagggc
     1321 cggtgtcgcc aggtggtgac tggcatggct tggctcttct tcgtatcatg gggtatgttc
     1381 cccatcctgt tcatcctcgg ccccgagggc ttcggcgtcc tgagccggta cggctccaac
     1441 gtcggccaca ccatcattga cctgatgtcg aagcagtgct ggggtctgct cggccactac
     1501 ctgcgcgtgc tgatccacag ccatatcctc atccacggcg acattcgcaa gaccaccaaa
     1561 ttgaacattg gtggcactga gattgaggtc gagacgctgg tggaggacga ggccgaggct
     1621 ggcgcggtcg cggcactcga gatggtgagc aagggcgagg agctgttcac cggggtggtg
     1681 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1741 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1801 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgat gtgcttcgcc
     1861 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1921 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1981 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2041 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2101 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2161 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2221 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     2281 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2341 atggacgagc tgtacaagta gaagcttatc gataatcaac ctctggatta caaaatttgt
     2401 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2461 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2521 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2581 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2641 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2701 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2761 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2821 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2881 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2941 tccctttggg ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct cataaagctc
     3001 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3061 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3121 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3181 ttgggaagac aatagcaggc atgctgggga tgcggtccgg actgtacaat tgcattcatt
     3241 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     3301 aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     3361 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3421 gctcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3481 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3541 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3601 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3661 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3721 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3781 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3841 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3901 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3961 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4021 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4081 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4141 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4201 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4261 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4321 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4381 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4441 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4501 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4561 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4621 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4681 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4741 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4801 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4861 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4921 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4981 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5041 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5101 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5161 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5221 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5281 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     5341 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5401 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5461 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5521 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5581 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5641 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5701 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5761 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     5821 gatcggtgcg ggcctcttcg ctattacgcc ag
//
