LOCUS       sequence-310681-        7185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes RsCCR1 rhodopsin domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7185)
  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 7185)
  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: RsCCR1_EYFP_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     615..634
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        615..633
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1689..1691,1692..2405)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1731..1752)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1992..2011)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2339..2360
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2435..2476
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2486..2503
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2526..2543)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2532..2756
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2802..3230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2889..2908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3099..3120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3239..3259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3244..3573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3424..3559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3486..3505
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3640..4434
                     /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(3694..3713)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4304..4323
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4610..4731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4647..4666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4701..4720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4780..4796)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4780..4796)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4793..4815)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4804..4820
                     /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(4828..4834,4835..4852,4853..4858))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4873..4894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5011..5028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5182..5770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5262..5281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5941..6732,6733..6801))
                     /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     6564..6583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6802..6906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6981..7000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7172..7185,1..366)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      601 tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta gttaagcttg
      661 gtaccgagct cggatccatg gtgctggacc tgcctgagga tttcgcccgg atgcacctgg
      721 agggcaccat ggagggctgg taccagccaa ccacatttgg cagcgccgca atcgtggcac
      781 actggatcac cttcctgttt ctgaccctgt gcacaatgta cctggccaga gactcctgga
      841 cagcaaaggg accaggaggc aaggagctgt acttcgccgg ctatcacgag gagtacaaca
      901 tctccctgta tgtgaatctg ttcgccatga tcgcctactt tggcaaggtg gtggccgatt
      961 ctatgggcca caactataac aatgtgggcc ccatgatcat cggcctgggc aattacagat
     1021 atgccgacta catgctgaca tgcccactgc tggcatggga tctgatggca caggtgcgcg
     1081 ccccttataa gatcaccggc gccatcctga tcttctgcgt gctgctgagc ggagcagcaa
     1141 caaactttta ccctggcgag cagatgaagc agggcgccat cgcctggttc tgctttggct
     1201 gtttcctgta tgctgccgcc tactatctgt tcttttctat cgtgaagaag cagtactcta
     1261 ggctggtggc cctgagcgcc aataccgagg ccaagaaggc cttcatgcca ctgaagctgg
     1321 ccctgtatac attctttagc atctgggtgg tgtttcccat cgtgtggatt ctgggctacc
     1381 acggcctgaa catcctgacc aatgacgccc aggagtgtct gcactgcgcc tgtgatctga
     1441 tcgccaagtc cttctatggc tttgccctgg ccaagtacag aacatatttc gacaagaaga
     1501 tgtacgatat gctggaggag ctgggagtgg acgcagagga gggcctggag cacctggaga
     1561 aggatctgaa ggaggtggac gatcccaaca atgtgaacaa tctgaagcgg ctgagcgtgg
     1621 cagcatccaa gaccagacag cacaaccacg ccaacatcaa tgccttttct gccgagcctg
     1681 cggccgccgt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1741 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1801 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1861 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     1921 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1981 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2041 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2101 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2161 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2221 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2281 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2341 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2401 agtaacatat gctctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     2461 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     2521 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2581 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2641 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2701 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2761 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2821 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2881 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2941 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3001 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3061 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3121 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3181 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3241 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3301 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3421 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3481 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3541 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3601 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3661 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3721 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3781 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3841 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3901 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3961 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4021 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4081 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4141 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4201 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4261 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4321 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4381 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4441 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4501 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4561 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4621 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4681 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4741 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4801 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4861 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4921 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4981 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5041 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5101 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5161 aaaggccgcg ttgctggcct ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5221 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5281 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5341 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5401 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5461 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5521 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5581 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5641 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5701 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5761 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5821 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5881 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5941 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6001 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6061 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6121 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6181 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6241 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6301 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6361 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6421 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6481 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6541 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6601 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6661 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6721 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6781 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6841 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6901 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6961 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7021 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7081 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     7141 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattg
//
