LOCUS       sequence-211926-        7038 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 7038)
  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 7038)
  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_653-EYFP
FEATURES             Location/Qualifiers
     source          1..7038
                     /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(1556..1558,1559..2272)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1598..1619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1859..1878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2206..2227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2302..2343
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2353..2370
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2393..2410)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2399..2623
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2669..3097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2756..2775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2966..2987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3106..3126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3111..3440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3291..3426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3353..3372
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3507..4301
                     /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(3561..3580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4171..4190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4477..4598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4514..4533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4568..4587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4647..4663)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4647..4663)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4660..4682)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4671..4687
                     /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(4695..4701,4702..4719,4720..4725))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4740..4761
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4878..4895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5049..5637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5129..5148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5808..6599,6600..6668))
                     /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     6431..6450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6669..6773)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6848..6867)
                     /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 catgccatac gataactatg gctggtggga
      721 gcgggagaat gccgactgca tctctaagat ggatgccaag ctggaccaga tgcacctggt
      781 gcagctggcc atgtccatgc tgtgcttttg tcagctggtg ttcatgacct ctagatttcc
      841 aaagcaggca tgggaggcaa tctacctgcc agcagcagag tgcgtgctgt atgcaatggc
      901 agcaagcgga aacggatacc tgaggctggc aaatggccgc ctgctgccat gggcaaggat
      961 ggcatcttgg ctggccacct gtcctgtgat gctgaaccag atcaatacaa tcagcccact
     1021 gaagctgtat ggcatcaacc tgaacaatgt gcagatcgcc ctgtccgtgg tgcgcgtggt
     1081 gaatggcgtg tctgccagct ccgtggacga tgacagcctg aagtggggct tctttttcat
     1141 gggcgtgttc tgcctggtca tcgagttttc catcgcctac aacatcttcg ccctgggcat
     1201 ggccaagttt aaggaggccg gcaaggattc tgacatctgc gcccgggcct gtgatagact
     1261 gcaggtgctg cggggcatct ttttcagcgc ctgggccatg ttccccatca tctggctgtt
     1321 ttcccctcag ggaacctgtc tgctgggaga ggatgtgatc cctctgctgt atctgttcgc
     1381 cgacctggtg tgcaagaatt gttttggcat cctgtggtac gataccacat ggaggctgct
     1441 ggacggcaag ggatggaacc ccgagctgca gctgacagac aatgaggtgg aggagctggt
     1501 gcacgccaag gaagaggtgg aggatcacca ccaccctcac gacgccgcgg ccgccgtgag
     1561 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1621 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1681 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1741 caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1801 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1861 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1921 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1981 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2041 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2101 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2161 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2221 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aacatatgct
     2281 ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     2341 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     2401 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2461 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2521 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2581 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2641 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2701 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2761 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2821 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2881 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2941 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3001 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3061 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3121 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3181 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3241 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3301 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3361 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3421 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3481 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3541 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3601 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3661 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3721 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3781 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3841 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3901 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3961 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4021 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4081 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4141 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4201 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4261 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4321 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4381 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4441 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4501 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4561 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4621 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4681 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4741 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4801 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4861 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4921 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4981 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5041 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5101 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5161 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5221 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5281 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5341 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5401 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5461 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5521 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5581 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5641 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5701 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5761 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5821 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5881 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5941 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6001 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6061 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6121 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6181 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6241 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6301 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6361 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6421 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6481 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6541 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6601 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6661 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6721 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6781 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     6841 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     6901 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     6961 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     7021 gggccagata tacgcgtt
//
