LOCUS       sequence-211927-        7074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anion channelrhodopsin from Rhodomonas sp. for expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7074)
  AUTHORS   Govorunova EG, Sineshchekov OA, Hemmati R, Janz R, Morelle O, 
            Melkonian M, Wong GK, Spudich JL
  TITLE     Extending the Time Domain of Neuronal Silencing with Cryptophyte 
            Anion Channelrhodopsins.
  JOURNAL   eNeuro. 2018 Jul 10;5(3). pii: eN-MNT-0174-18. doi: 
            10.1523/ENEURO.0174-18.2018. eCollection 2018 May-Jun.
  PUBMED    30027111
REFERENCE   2  (bases 1 to 7074)
  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: "eNeuro."; 
            date: "2018-07-10"; volume: "5(3). pii: eN-MNT-0174-18. doi"; pages:
            " 10.1523/ENEURO.0174-18.2018. eCollection 2018 May-Jun"
COMMENT     Sequence Label: pcDNA3.1-R1ACR_6367-EYFP
FEATURES             Location/Qualifiers
     source          1..7074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1592..1594,1595..2308)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1634..1655)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1895..1914)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2242..2263
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2338..2379
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2389..2406
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2429..2446)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2435..2659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2705..3133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2792..2811)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3002..3023
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3142..3162)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3147..3476
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3327..3462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3389..3408
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3543..4337
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3597..3616)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4207..4226
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4513..4634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4550..4569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4604..4623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4683..4699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4683..4699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4696..4718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4707..4723
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4731..4737,4738..4755,4756..4761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4776..4797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4914..4931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5085..5673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5165..5184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5844..6635,6636..6704))
                     /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     6467..6486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6705..6809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6884..6903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc catgtctgcc atcacagacc ccgccctggg
      721 cctgtacgga acctgggagg ccgccaacaa ggagtgcgtg gataagggcg tggagtttag
      781 ctccaatacc cggatgatcg tgctgctgtt tgccagcgtg tgcttctgtc agacagtgta
      841 tatgaccagc agggcaccaa gggtgaactg ggagggctgc tacctgcctc tgtgcgagac
      901 atttctgtat ggcctggcag caagcggaaa cggaatgatc cggctgtcca atggcagaat
      961 ggtgccaatg gccaggttct gctcctggct ggtgacctgt cccatcatgc tgatgtctgt
     1021 gctggccatc cacgacgtgc gctggatggg cgtgcctgtg aagaatatgg tcatcgcagc
     1081 cacactgatc aggaccgtgc tgggaatcgg agcatctagc accatcacag agacactgag
     1141 atgggtgcag tactgcatcg cctgtgccgt gttcgtgttc gagctgacaa tggtgaacca
     1201 gctgtttatc gccggcctgc agaagttcgg cgtggtgaag accaagctga atgacatggt
     1261 gtatatgagg atcatgatcc tgcgcgccct gttctttttc agctggacag cctttcctgt
     1321 gatctggatt ctgtcctcta ccggcttctg cgtgatcgga gaggacggct ccgtgctggc
     1381 atacctggtg gccgacttcc tgtgcaagaa cgtgtatggc atcatcaaca caaataccct
     1441 gttccaggtg ctggatggcg agtggaagcc agagacaagg gacaccgatg gcaagatcgc
     1501 cacaaatcgc cacgcccaca tcggcgagac atacctggag atgctgctgg cccggatcag
     1561 acggcacaga gagggcgagc tggcggccgc cgtgagcaag ggcgaggagc tgttcaccgg
     1621 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1681 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1741 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg
     1801 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1861 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1921 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1981 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2041 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2101 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2161 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     2221 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2281 tctcggcatg gacgagctgt acaagtaaca tatgctctag agggcccgcg gttcgaaggt
     2341 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat
     2401 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2461 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2521 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2581 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2641 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2701 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2761 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2821 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2881 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2941 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3001 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3061 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3121 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3241 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3301 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3361 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3421 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3481 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3541 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3601 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3661 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3721 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3781 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3841 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3901 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3961 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4021 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4081 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4141 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4201 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4261 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4321 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     4381 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4441 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4501 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4561 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4621 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4681 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4741 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4801 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4861 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4921 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4981 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5041 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5101 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5161 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5221 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5281 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5341 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5401 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5461 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5521 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5581 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5641 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5701 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5761 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5821 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5881 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5941 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6001 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6061 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6121 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6181 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6241 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6301 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6361 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6421 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6481 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6541 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6601 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6661 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6721 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6781 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6841 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6901 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6961 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7021 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
