LOCUS       sequence-310661-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes GtCCR2 rhodopsin domain .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Sineshchekov OA, Govorunova EG, Li H, Spudich JL
  TITLE     Bacteriorhodopsin-like channelrhodopsins: Alternative mechanism for 
            control of cation conductance.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 7;114(45):E9512-E9519. doi: 
            10.1073/pnas.1710702114. Epub 2017 Oct 25.
  PUBMED    29078348
REFERENCE   2  (bases 1 to 7083)
  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"; doi: 
            "10.1073/pnas.1710702114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-11-7- 7"; volume: "114"; issue: "45"; pages: 
            "E9512-E9519"
COMMENT     Sequence Label: GtCCR2_EYFP_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        39..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     667..686
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        667..685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      728..737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1639..1641,1642..2355)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1681..1702)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1942..1961)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2289..2310
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2385..2426
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2436..2453
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2476..2493)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2482..2706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2752..3180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2839..2858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3049..3070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3189..3209)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3194..3523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3374..3509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3436..3455
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3590..4384
                     /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(3644..3663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4254..4273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4560..4681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4597..4616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4651..4670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4730..4746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4730..4746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4743..4765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4754..4770
                     /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(4778..4784,4785..4802,4803..4808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4823..4844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4961..4978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5132..5720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5212..5231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5891..6682,6683..6751))
                     /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     6514..6533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6752..6856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6931..6950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      661 cgaaattaat acgactcact atagggagac ccaagctggc tagttaagct tggtaccgag
      721 ctcggatcca tggtggccag cagcgccgtg attaccgcca actggatcag ctttctggcc
      781 atcagcgcca gcttcatcat cctgctggtc atctccctga ggtacaaagg ccccggcgga
      841 accgagagct tctacaacgg cttcaaggag cagaacatgc tgaccgtgtt catcaatctg
      901 tggtgtgccc tggcctactt tgccaaagtg ctgcagtccc actccaacga taatggcttc
      961 gcccctctga ccgtgatccc ttacgtggac tactgcacca cctgccctct cctgacactg
     1021 gatctgctgt ggtgcctgga cgccccttac aaaatcagca gcgccgtgct cgtgttcacc
     1081 tgcctggtga ttgccgtcgc ttgcagcctg gccgtggccc ctttcagcta ctgctggttt
     1141 gccatgggaa tggtgctctt caccttcacc tacgtgttca tcctgtccat cgtgaggcag
     1201 agactggact tcttcacact gtgcgccaga gacagcaacg ccaaacagag cctgaagcat
     1261 ctgaagaccg ccgtgttcat ctacttcggc atctggctgc tgttccccct cctgtggctg
     1321 ctctcctata gggccgccaa cgtgatcagc aatgacatca accacatctt ccactgcatc
     1381 ctggacgtga tcgccaagag cgtgtacggc tttgccctcc tgtactttaa gatgtacttt
     1441 gacaaaaagc tgatcgagag cggcatcgac gaggacgact tcgccaagtt ctccaaggtc
     1501 gtgaccaccc acagaagcga ggagaagcag aagaggaagc ccgccgtgtc ccaaagccct
     1561 aagatgtact acgacgaggc cgcttaccag gacggcgagg tcgagagcaa cctgcagtcc
     1621 aagatcagag cggccgccgt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1681 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1741 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1801 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
     1861 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1921 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1981 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2041 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2101 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2161 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2221 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     2281 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2341 gagctgtaca agtaacatat gctctagagg gcccgcggtt cgaaggtaag cctatcccta
     2401 accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt
     2461 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2521 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2581 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2641 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2701 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2761 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2821 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2881 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2941 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3001 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3061 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3121 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3181 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3241 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3301 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3361 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3421 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3481 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3541 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3601 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3661 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3721 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3781 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3841 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3901 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3961 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4021 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4081 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4141 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4201 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4261 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4321 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4381 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4441 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4501 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4561 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4621 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4681 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4741 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4801 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4861 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4921 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4981 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5041 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5101 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5161 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5221 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5281 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5341 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5401 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5461 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5521 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5581 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5641 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5701 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5761 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5821 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5881 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5941 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6001 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6061 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6121 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6181 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6241 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6301 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6361 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6421 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6481 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6541 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6601 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6661 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6721 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6781 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6841 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6901 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6961 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7021 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     7081 ttt
//
