LOCUS       sequence-211928-        7119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Anion channelrhodopsin from Rhodomonas lens for expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7119)
  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 7119)
  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-RlACR_497-EYFP
FEATURES             Location/Qualifiers
     source          1..7119
                     /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(1637..1639,1640..2353)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1679..1700)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1940..1959)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2287..2308
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2383..2424
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2434..2451
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2474..2491)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2480..2704
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2750..3178
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2837..2856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3047..3068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3187..3207)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3192..3521
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3372..3507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3434..3453
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3588..4382
                     /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(3642..3661)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4252..4271
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4558..4679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4595..4614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4649..4668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4728..4744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4728..4744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4741..4763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4752..4768
                     /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(4776..4782,4783..4800,4801..4806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4821..4842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4959..4976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5130..5718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5210..5229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5889..6680,6681..6749))
                     /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     6512..6531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6750..6854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6929..6948)
                     /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 catggagtac cagagcctgg acgattttcc
      721 actggagaca ctgtacttcc ggaacggcac agaggtggac ccctatgatc acagatgcaa
      781 gctgggcatc cctggaggct cccagggcat ccacaccttt cagctggtgt gggcactgct
      841 gtgcgcagca caggccctgt ttctgttctc caagtggcca aaagtgggct gggaggccat
      901 gtacatgccc tgctgtgagg ccctgtctta tggcctgctg agctccggct acggctatat
      961 gaagctgcac ggcgagttcc actacctgcc ttggatcagg tacgccgcct ggatcgtgac
     1021 cacaccagtg ctgctgaacc aggtgcaggg catgatcgac atcaagatcg gctctctgag
     1081 gctggcacca atccagagcc tggcaaacct gatcatgatc atcatgggcg tgagcgccca
     1141 cctgcaggag aatcacgcct ggaagtggct gttctttttc gtgggctgcg tgatgtgcgc
     1201 cgtgatctac tatatcgcct ggctgcagtt tgccaaagtg aatgagctgt acgccgccac
     1261 caagacagag atcggcgata aggtgtgctt tcagctgaag ctgatcatgt tttgtttcta
     1321 tgtgggctgg tccatgttct ctgtgatctg gctgctgtct agcgagggcc tgtgcctgat
     1381 cgaggagcac atcagcgccg ccgcccacat cccaggcgac atcctgagca agaactcctt
     1441 tggcctgctg atgtggtata ccaggtggat cacactggga ggagcatggg atatcaataa
     1501 gttcctggag aagcaggcca aggagaaggc cgaggccgtg ctgacacctg ccgagaagag
     1561 gaaggccatc atgaagcagg cagaggcatg cttctccatg gcagacgcct gtctgcagca
     1621 ggccaatgcg gccgccgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1681 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1741 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1801 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
     1861 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1921 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1981 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2041 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2101 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2161 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2221 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     2281 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2341 gctgtacaag taacatatgc tctagagggc ccgcggttcg aaggtaagcc tatccctaac
     2401 cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca ttgagtttaa
     2461 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2521 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2581 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2641 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2701 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     2761 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2821 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2881 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2941 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3001 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3061 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3121 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3181 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3241 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3301 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3361 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3421 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3481 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3541 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3601 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3661 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3721 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3781 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3841 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3901 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3961 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4021 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4081 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4141 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4201 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4261 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4321 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4381 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4441 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4501 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4561 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4621 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4681 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4741 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4801 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4861 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4921 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4981 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5041 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5821 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5881 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5941 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6001 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6061 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6121 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6181 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6241 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6301 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6361 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6421 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6481 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6541 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6601 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6661 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6721 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6781 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6841 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     6901 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6961 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7021 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     7081 ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
