LOCUS       sequence-310683-        7110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes RsCCR2 rhodopsin domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7110)
  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 7110)
  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: RsCCR2_EYFP_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     695..715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     789..808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        789..807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1788..1790,1791..2504)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1830..1851)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2091..2110)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2438..2459
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2534..2575
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2585..2602
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2625..2642)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2631..2855
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2901..3329
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2988..3007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3198..3219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3338..3358)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3343..3672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3523..3658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3585..3604
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3739..4533
                     /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(3793..3812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4403..4422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4709..4830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4746..4765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4800..4819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4879..4895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4879..4895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4892..4914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4903..4919
                     /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(4927..4933,4934..4951,4952..4957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4972..4993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5110..5127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6040..6831,6832..6900))
                     /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     6663..6682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6901..7005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7080..7099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
       61 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      121 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      661 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      721 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      781 atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag cttggtaccg
      841 agctcggatc catggccgcc ggcaacgagc acttcgagac accacaggag gcccggatgg
      901 tgctgatcgc cgactacgtg gccttttgct gtctggccgc ctgcgccttc tttctgcacc
      961 ggagaatcgt gatgttccgg ggccccacag gcgacgatca ctactttatg ggctatagag
     1021 aggagaagat gctgtgcgtg ctgatcacca tcttcgccgc catcggctat tggagccgga
     1081 tcacagccag agtgcagggc agccacaagg ccctggacct gtccaccctg cagtacgtgg
     1141 attatctggc cacctgccct ctggtgacag tggacatgct gtggtccctg aatctgccat
     1201 acaagtttac atggggcgcc ctggtgttcc tgtgcatcct gtctgccttt gcctgtgatg
     1261 caatggaggg acaggcaagg tggatgtggt tcggcgtggg catgggcctg ttttcttacg
     1321 tgtggatcag cgtgggcagg ctggtgaagt tccgcctgac ccagctggcc gccctgagag
     1381 cagagggcgt gctgaaggtg gatatcgagt accccctgaa ggtcggcacc gccacattct
     1441 ttttcctgtg gttcgcctat cctctgatgt ggctgctgga ggagtttgac gtgatcaacc
     1501 acgccaagtc tgaggtggcc cacgtggtgt ttgatgtggt ggtgaagagc gtgtacgcca
     1561 tctgcctgca ggagtttggc ttcaggtgtg accgcaccta ttttgagttc cccagcatgt
     1621 ccgtgaccgt ggtgcctgat gaggagtacc agacaaagaa cgagcacgcc atggacgagt
     1681 atgccaatga gaggctgcag gatgcctggc acaagctgcc agtgaatgag acacagcgcc
     1741 acacagtgcc caagtcctgg cactgtcagg gctctatggc ggccgccgtg agcaagggcg
     1801 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1861 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1921 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     1981 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2041 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2101 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2161 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2221 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2281 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2341 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     2401 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2461 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaacatatg ctctagaggg
     2521 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     2581 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     2641 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2701 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2761 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2821 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2881 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3241 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3301 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3361 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3421 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3481 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3541 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3601 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3661 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3721 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3781 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3841 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3901 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3961 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4021 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4081 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4141 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4201 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4261 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4321 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4381 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4441 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4501 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     4561 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4681 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4741 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4801 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4861 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcctt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7021 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7081 acaatctgct ctgatgccgc atagttaagc
//
