LOCUS       Exported                5609 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Anion-conducting Channelrhodopsin with near-complete desensitization
            in continuous light. Codon-optimized for mammalian expression 
            (human/mouse)..
ACCESSION   .
VERSION     .
KEYWORDS    p-mCherry-C1-MerMAID2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5609)
  AUTHORS   Oppermann J, Fischer P, Silapetere A, Liepe B, Rodriguez-Rozada S, 
            Flores-Uribe J, Peter E, Keidel A, Vierock J, Kaufmann J, Broser M, 
            Luck M, Bartl F, Hildebrandt P, Simon Wiegert J, Beja O, Hegemann P,
            Wietek J
  TITLE     MerMAIDs: a family of metagenomically discovered marine 
            anion-conducting and intensely desensitizing channelrhodopsins.
  JOURNAL   Nat Commun. 2019 Jul 25;10(1):3315. doi: 10.1038/s41467-019-11322-6.
  PUBMED    31346176
REFERENCE   2  (bases 1 to 5609)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5609
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        45..348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        349..552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     503..523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      579..588
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1500..2207
                     /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(1642..1660)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1910..1929
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2381..2502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2418..2437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2472..2491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2509..2964)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2646..2667)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2858..2877
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2991..3095
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3097..3454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3120..3140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3305..3440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3367..3386
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3489..4283
                     /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(3543..3562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4153..4172
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4471..4490)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4515..4562
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4891..5479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5380..5399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
       61 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      121 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      181 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      241 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      301 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      361 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      421 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      481 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      541 ataagcagag ctggtttagt gaaccgtcag atccgctagc caccatggtc tcagtccagc
      601 tgcatgtctt cgctcagatt ctgactacct tcatcagttg tgctaacgca atgtactacc
      661 tgtatatcta taagtgcacc aagaggggcc acctgatgaa gtgggaggtg gcctgggtgg
      721 ccatcgtgga gaccatcaac tacacaatcc agatcagcac cggctccgag cagatcgagc
      781 tggccggagg agcaaagttc ccatggatga gatatgtggg ctggcagctg acatgtcccg
      841 tgctgctgac ctttatcgtg acaaatatgc tggaggcacc aaccacacgc gtggtggtgc
      901 agatgctgat gctgctgcag ttcatcatgc tgtccggcat gaccgcctct ctgctgcagg
      961 ccatgaccct gaaagtgatc tttgtgctgc tggccagcgc cggactgatc tgcctgtact
     1021 attccctgta taccaagcgg aagaacaagc agctgcccat ggccaacaat aaggagatgc
     1081 acctgctgct gtatttcatg acatcctggc tggtgttccc actgatgttt atcctgggcc
     1141 ccgagatgct gaacgtgatg cctttcgagt ggaccctggt gggccactgt atcggcgacc
     1201 tgatctctaa gaatgccttt ggcatgctgg cctggcagta cacaaagtat ctgcacaggg
     1261 atgagatcaa ggagatcaag agaggcttcc ctgaggtgga cgatgtggag tctatccaca
     1321 gcgccgcagc cgtgtctacc ggccagggca agaccgccac acacatcagc ctgcccctga
     1381 acatgctgcc tatgttcatg tataacaacc agaacacacc aaagcacgac aatcccagca
     1441 ctccaaagca tcatactcca gaagaaatca ctgacgccgc aactaaggct ggaccggtgg
     1501 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     1561 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1621 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     1681 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     1741 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     1801 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     1861 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     1921 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1981 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     2041 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     2101 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     2161 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagag ctcaagcttc
     2221 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat
     2281 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2341 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2401 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2461 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     2521 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2581 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2641 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2701 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2761 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2821 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2881 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2941 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3001 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3061 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3121 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3181 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3301 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3361 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3421 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3481 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3541 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3601 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3661 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3721 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3781 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3841 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3901 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3961 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4021 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4081 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4141 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4201 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4261 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4321 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4381 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4441 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4501 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4561 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4621 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4681 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4741 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4801 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4861 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4921 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4981 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5041 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5101 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5161 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5221 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5281 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5341 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5401 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5461 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5521 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5581 taccgccatg cattagttat taatagtaa
//
