LOCUS       Exported                5635 bp ds-DNA     circular SYN 12-MAY-2021
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    p-mCherry-C1-Phobos_CA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5635)
  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 5635)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5635
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        130..433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        434..637
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     588..608
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      666..675
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1611..2318
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1753..1771)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2021..2040
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2492..2613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2529..2548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2583..2602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2620..3075)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2757..2778)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2969..2988
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3102..3206
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3208..3565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3231..3251)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3416..3551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3478..3497
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3600..4394
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3654..3673)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4264..4283
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4582..4601)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4626..4673
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5002..5590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5491..5510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
       61 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      121 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      181 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      241 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      301 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      361 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      421 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      481 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      541 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      601 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      661 ctagcgccac catggactat ggcggcgctt tgtctgccgt cggactcttc caaacctcat
      721 acactcttga gaacaatggt tctgttattt gcatcccgaa caacggccag tgcttctgct
      781 tggcttggct taaatccaac ggaacaaatg ccgagaagtt ggctgccaac attctgcagt
      841 ggatttcttt tgcgctttca gcgctctgcc tgatgttcta cggctaccag acctggaagt
      901 ctacttgcgg ctgggagaac atttacgtgg ccacgatcca gatgatcaag ttcatcatcg
      961 agtatttcca tagctttgac gaacctgcgg tgatctactc atccaacggc aacaagaccc
     1021 ggtggcttcg ttacgcgagc tggctgctga ccgcccctgt cattcttatc cacctgagca
     1081 accttacggg tctggcgaac gactataaca agcgtaccat gggtctgctg gtgtcagata
     1141 tcggcggcat cgtgtgggcc accacggccg cgctgtccaa gggatacgtc cgtgtcattt
     1201 tcttcctgat gggcctgtgc tacggcatct acacattctt caacgcagcc aaggtctaca
     1261 ttgaggcgta ccacaccgtg ccgaagggcc ggtgtcgcca ggtggtgact ggcatggctt
     1321 ggctcttctt cgtatcatgg ggtatgttcc ccatcctgtt catcctcggc cccgagggct
     1381 tcggcgtcct gagccggtac ggctccaacg tcggccacac catcattgac ctgatgtcga
     1441 agcagtgctg gggtctgctc ggccactacc tgcgcgtgct gatccacagc catatcctca
     1501 tccacggcga cattcgcaag accaccaaat tgaacattgg tggcactgag attgaggtcg
     1561 agacgctggt ggaggacgag gccgaggctg gcgcggtcgc ggcaccggtg gtgagcaagg
     1621 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     1681 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1741 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     1801 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     1861 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1921 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     1981 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2041 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     2101 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     2161 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     2221 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2281 gccactccac cggcggcatg gacgagctgt acaagtaaga gctcaagctt cgaattctgc
     2341 agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca
     2401 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2461 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2521 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2581 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     2641 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2701 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2761 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2821 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     2881 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     2941 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3001 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3061 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3121 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3181 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3241 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3301 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3361 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3421 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3481 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3541 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3601 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3661 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3721 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3781 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3841 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3901 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3961 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4021 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4081 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4141 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4201 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4261 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4321 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4381 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4441 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4501 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4561 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4621 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     4681 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4741 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4801 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     4861 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4921 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4981 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5041 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5101 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5161 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5221 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5281 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5341 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5401 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5461 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5521 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5581 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctgg
//
