LOCUS       sequence-310678-        7131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes RaCCR2 rhodopsin domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7131)
  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 7131)
  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: RaCCR2_EYFP_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        126..505
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        506..709
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     660..680
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     754..773
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        754..772
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1774..1776,1777..2490)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1816..1837)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2077..2096)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2520..2561
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2571..2588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2611..2628)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2617..2841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2887..3315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2974..2993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3184..3205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3324..3344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3329..3658
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3509..3644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3571..3590
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3725..4519
                     /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(3779..3798)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4389..4408
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4695..4816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4732..4751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4786..4805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4865..4881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4865..4881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4878..4900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4889..4905
                     /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(4913..4919,4920..4937,4938..4943))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4958..4979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5096..5113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5267..5855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5347..5366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6026..6817,6818..6886))
                     /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     6649..6668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6887..6991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7066..7085)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
       61 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      121 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      181 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      241 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      301 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      361 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      421 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      481 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      541 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      601 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      661 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      721 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      781 agctggctag ttaagcttgg taccgagctc ggatccatgc caatgttcgg cggccggaga
      841 agcgagtccg agaaggcccc ccctatcgag ggcgatgagc ggaacatcct gatcgccgac
      901 atcgtgggct tctcctgcct gggcatcacc gcctttatcc tgaattacag gctgaaggcc
      961 ttcaagggac caagggacga tgacaagtac tttgtgggct atcgggagga gaagatgctg
     1021 gccctgctga tcaacctgtt tgccacagtg ggctactggg ccaagatctg cgccagatat
     1081 gattctaata tgggcagctc cgtggacatc accacatacg agtatctgga ttacctggcc
     1141 acctgtcctc tggtgacact ggacatgctg tggaccctga acctgccata taagtttaca
     1201 tccggcgcca tgatcttcct gtgcatcctg tgcgcctttg cctctgagat caccgccggc
     1261 acagccaggt ggatgtggtt cggctttggc ttcatcctgt tcctgtacgt gtggaccaca
     1321 gtgggcagcc tggtgaagat gcgcctgcag cagctgaaga ccctgacaga tcagggcgtg
     1381 ctggtgccag gacagacctc cctgaagatc gccaccgcca cattcttttg cctgtggctg
     1441 ggctaccctc tgctgtggct gatcctggac ctgggcggca tgacagagtt ccaggccgag
     1501 atctgtcacg tgggctttga tgtggtggtg aagtctctgt atggcctggg catgctggag
     1561 ttcgtgatgc aggccgacag aatccacttt gagttcgaga cactgtctcc acccgatgac
     1621 tacggcagcg agtctagcaa gaattatggc aaggacctgg agagcggcca gcagccttcc
     1681 tcttttgata ccttcaaggg cacacacgcc gccaaggagg agaaggagga gaaggaggat
     1741 gaggaggacg cccacaagac cgatgcggcc gccgtgagca agggcgagga gctgttcacc
     1801 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1861 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1921 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag
     1981 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2041 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2101 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2161 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2221 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2281 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2341 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     2401 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2461 actctcggca tggacgagct gtacaagtaa catatgctct agagggcccg cggttcgaag
     2521 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc
     2581 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2641 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2701 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2761 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2821 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2881 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2941 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3061 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3181 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3241 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3301 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3481 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3661 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3721 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3781 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3841 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3901 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3961 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4021 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4081 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4141 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4201 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4261 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4321 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4381 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4441 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4501 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     4561 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4621 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4681 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4741 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4801 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4861 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4921 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4981 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcctttttc cataggctcc
     5281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6001 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6061 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7021 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     7081 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg c
//
