LOCUS       sequence-310663-        7176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes RaCCR1 rhodopsin domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7176)
  AUTHORS   Sineshchekov OA, Govorunova EG, Li H, Wang Y, Melkonian M, Wong GK, 
            Brown LS, Spudich JL
  TITLE     Conductance Mechanisms of Rapidly Desensitizing Cation 
            Channelrhodopsins from Cryptophyte Algae.
  JOURNAL   mBio. 2020 Apr 21;11(2). pii: mBio.00657-20. doi: 
            10.1128/mBio.00657-20.
  PUBMED    32317325
REFERENCE   2  (bases 1 to 7176)
  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: "mBio."; 
            date: "2020-04-21"; volume: "11(2). pii: mBio.00657-20. doi"; pages:
            " 10.1128/mBio.00657-20"
COMMENT     Sequence Label: RaCCR1_EYFP_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        159..538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        539..742
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     693..713
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     787..806
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        787..805
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1852..1854,1855..2568)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1894..1915)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2155..2174)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2502..2523
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2598..2639
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2649..2666
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2689..2706)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2695..2919
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2965..3393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3052..3071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3262..3283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3402..3422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3407..3736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3587..3722
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3649..3668
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3803..4597
                     /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(3857..3876)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4467..4486
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4773..4894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4810..4829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4864..4883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4943..4959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4943..4959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4956..4978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4967..4983
                     /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(4991..4997,4998..5015,5016..5021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5036..5057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5174..5191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5345..5933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5425..5444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6104..6895,6896..6964))
                     /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     6727..6746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6965..7069)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7144..7163)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
       61 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      121 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      181 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      241 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      301 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      361 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      421 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      481 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      541 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      601 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      661 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      721 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      781 cgaaattaat acgactcact atagggagac ccaagctggc tagttaagct tggtaccgag
      841 ctcggatcca tggtgtctga gggctacggc gacaagtatc tggagggcag catgaggggc
      901 tggtacacac cctgcacctt cggcacacct gccatcatgg cccactgggt gaccttcctg
      961 tttctgttcg tgtgcacaat gtatctggca agggacagct ggaccgcaag aggaccaggc
     1021 ggcagagaga gatggtacgc cggctatcac gaggattaca acatcagcct gtatgtgaat
     1081 ctgtttgcca tgatcgccta cttcggcaag atcgtggccg actccgtggg ccacaactat
     1141 aacaatgtgg gccccctggt catcggcctg ggcaattaca gatatgccga ctacatgctg
     1201 acctgccctc tgctggccta tgatctgatg tcccaggtgc gcgccccctt caagatcaca
     1261 ggcggcgtgc tgatcttctg cgtgctgctg accggcacag ccaccaacta ctatcctggc
     1321 gatgagatgc gggccggctc tatggcctgg tttatcttcg gctgctttct gtacttcgtg
     1381 gcctactatc tgttttacac cgtggtgaag atccagtata caagactgct ggccctggca
     1441 gcatctaccg aggccaagaa ggcctttatg cccctgaagc tggccctgta cacattcttt
     1501 agcatctgga tcgtgttccc tatcgtgtgg ctgctgggcc accagggcct gaacatcctg
     1561 tccaatgacg cccaggagtg tctgcactgc gcctgtgatc tgaccgccaa gtctttttac
     1621 ggcttcgccc tggccaagta ccggaagtat tttgacaaga agatgttcga tatgctgaca
     1681 gagctgggaa tcgacggaga ggagggcctg gagcacctgg agcaggatct gaagaaccag
     1741 gacatcggcg atggcaaggc cgatatcaag aggatgtcca ccacaagggg accaggcctg
     1801 agcgtgatct ccgtgaataa gcaccacccc gccttcctgg ccgcggccgc cgtgagcaag
     1861 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1921 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1981 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2041 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2101 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2161 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2221 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2281 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2341 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2401 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2461 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2521 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaca tatgctctag
     2581 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     2641 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     2701 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2761 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2821 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2881 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2941 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3001 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3061 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3121 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3181 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3241 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3301 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3361 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3481 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3541 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3601 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3661 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3721 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3781 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3841 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3901 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3961 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4021 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4081 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4141 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4201 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4261 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4321 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4381 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4441 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4501 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4561 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     4621 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4681 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4741 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4801 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4861 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4921 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4981 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5041 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5101 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5341 cctttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6061 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7081 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7141 cagtacaatc tgctctgatg ccgcatagtt aagcca
//
