LOCUS       sequence-256051-        6022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Slow cycling (open for minutes) step-function artificial anion 
            conducting channelrhodopsin (aACR). High light sensitivity. 
            Activation with blue light, inactivation max with 580 nm 
            (accelarates closure to ms). Codon optimized for mammalian 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6022)
  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 6022)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2017-11-2"; volume: "7(1):14957. doi"; pages: " 
            10.1038/s41598-017-14330-y"
COMMENT     Sequence Label: AAV2_hSyn_Phobos_CA_Citrine
FEATURES             Location/Qualifiers
     source          1..6022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        226..673
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      693..702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1638..1640,1641..1643,1644..2354)
                     /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(1683..1704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1944..1963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2291..2312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2370..2958
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2423..2443)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2841..2852)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2999..3206
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3376..3505
                     /label=AAV2 ITR
     rep_origin      complement(3742..4330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3822..3841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4501..5292,5293..5361))
                     /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     5124..5143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5362..5466)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5492..5947)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5629..5650)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5841..5860
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(6019..6022,1..126)
                     /label=AAV2 ITR
ORIGIN
        1 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
       61 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga
      181 agatcgtagg ttaattaatc tagactgcag agggccctgc gtatgagtgc aagtgggttt
      241 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      301 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      361 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      421 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      481 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      541 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      601 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      661 cctgagagcg cagtcgaatt caagctgcta gcgccaccat ggactatggc ggcgctttgt
      721 ctgccgtcgg actcttccaa acctcataca ctcttgagaa caatggttct gttatttgca
      781 tcccgaacaa cggccagtgc ttctgcttgg cttggcttaa atccaacgga acaaatgccg
      841 agaagttggc tgccaacatt ctgcagtgga tttcttttgc gctttcagcg ctctgcctga
      901 tgttctacgg ctaccagacc tggaagtcta cttgcggctg ggagaacatt tacgtggcca
      961 cgatccagat gatcaagttc atcatcgagt atttccatag ctttgacgaa cctgcggtga
     1021 tctactcatc caacggcaac aagacccggt ggcttcgtta cgcgagctgg ctgctgaccg
     1081 cccctgtcat tcttatccac ctgagcaacc ttacgggtct ggcgaacgac tataacaagc
     1141 gtaccatggg tctgctggtg tcagatatcg gcggcatcgt gtgggccacc acggccgcgc
     1201 tgtccaaggg atacgtccgt gtcattttct tcctgatggg cctgtgctac ggcatctaca
     1261 cattcttcaa cgcagccaag gtctacattg aggcgtacca caccgtgccg aagggccggt
     1321 gtcgccaggt ggtgactggc atggcttggc tcttcttcgt atcatggggt atgttcccca
     1381 tcctgttcat cctcggcccc gagggcttcg gcgtcctgag ccggtacggc tccaacgtcg
     1441 gccacaccat cattgacctg atgtcgaagc agtgctgggg tctgctcggc cactacctgc
     1501 gcgtgctgat ccacagccat atcctcatcc acggcgacat tcgcaagacc accaaattga
     1561 acattggtgg cactgagatt gaggtcgaga cgctggtgga ggacgaggcc gaggctggcg
     1621 cggtcgcggc actcgagatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1681 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1741 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1801 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct
     1861 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1921 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1981 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2041 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     2101 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2161 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2221 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     2281 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2341 acgagctgta caagtagaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2401 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2461 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2521 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2581 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2641 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2701 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2761 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2821 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2881 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2941 tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgctcata aagctcgact
     3001 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3061 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3121 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3181 gaagacaata gcaggcatgc tggggatgcg gtccggactg tacaattgca ttcattttat
     3241 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3301 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     3361 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3421 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3481 cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg
     3541 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3601 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3661 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3721 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3781 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3841 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3901 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3961 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4021 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4081 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4141 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4201 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4261 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4321 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4381 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4441 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4501 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4561 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4621 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4681 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4741 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4801 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4861 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4921 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4981 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5041 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5101 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5161 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5221 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5281 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5341 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5401 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5461 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5521 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5581 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5641 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5701 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5761 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5821 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5881 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5941 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6001 tcttcgctat tacgccagct gc
//
