LOCUS       sequence-310659-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes GtCCR1 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: GtCCR1_EYFP_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..477
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        478..681
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     632..652
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..745
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        726..744
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1698..1700,1701..2414)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1740..1761)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2001..2020)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2348..2369
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2444..2485
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2495..2512
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2535..2552)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2541..2765
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2811..3239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2898..2917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3108..3129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3248..3268)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3253..3582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3433..3568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3495..3514
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3649..4443
                     /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(3703..3722)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4313..4332
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4619..4740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4656..4675)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4710..4729
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4789..4805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4789..4805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4802..4824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4813..4829
                     /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(4837..4843,4844..4861,4862..4867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4882..4903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5020..5037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5191..5779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5271..5290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5950..6741,6742..6810))
                     /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     6573..6592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6811..6915)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6990..7009)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
       61 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
      121 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      181 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      241 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      301 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      361 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      421 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      481 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      541 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      601 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      661 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
      721 gaaattaata cgactcacta tagggagacc caagctggct agttaagctt ggtaccgagc
      781 tcggatccat ggtcgagagc tccgccgtca tcgccgccaa ctggatcagc tttctggtga
      841 tcgccggcag ctttgtcgtg ctgtgcttca tctccctgag atacaagggc cccggaggca
      901 atgagaacta ctacaatggc ttcagggagc agaacatgct gacagtgatc atcaacctgt
      961 ggtgcgccct ggcctacttc gccaaggtgc tgcagagcca cagcgacgac gatggcttcg
     1021 tgcccctcac caagatccct tacctggact acgccaccac atgccctctg ctgaccctgg
     1081 acctgatgtg gtgcctcgac gccccttaca agatcacatc cgccgtcctg gtgttcaccg
     1141 tgatgatcac aggagtggcc tgttccctgg ccgtggcccc ttacagcttt tactggttcg
     1201 ccatgggcat ggtcctgttc atcttcacat acgtgctcat gctgagcatc gtcagagaga
     1261 ggctggagtt cattacccag tgtgcccacg actccaatgc caaaaggtcc atcaagcacc
     1321 tgaaggccgc cgtgatcatc tacttcggca tctggcccat cttcgccatc ctctggctgc
     1381 tgagctacag agccgccaac gtgatcagca acgacacaaa ccacatcctg cactgcatcc
     1441 tggacgtgat cgctaaaagc tgcttcggct tcgtgctgct gcacttcaag atgtacttcg
     1501 acaagaagct gctggagagc ggagtgcatg aggatgagtt tgccgagtac tccaaggagg
     1561 tgtccaagca cagagaggag aagccctgga ggaagaccgc cagcagctac cctgagtccg
     1621 actacgacga gcctgtgtac atgaacggcg aggacctgaa cctgaacgac ccccccgctt
     1681 ggggcggcgc ggccgccgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1741 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1801 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1861 tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc
     1921 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1981 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2041 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2101 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2161 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2221 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2281 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     2341 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2401 agctgtacaa gtaacatatg ctctagaggg cccgcggttc gaaggtaagc ctatccctaa
     2461 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta
     2521 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2581 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2641 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2701 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2761 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2821 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2881 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2941 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3001 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3061 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3121 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3181 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3241 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3301 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3361 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3421 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3481 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3541 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3601 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3661 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3721 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3781 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3841 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3901 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3961 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4021 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4081 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4141 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4201 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4261 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4321 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4381 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4441 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4501 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4561 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4621 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4681 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4741 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4801 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4861 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4921 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4981 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5041 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5101 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5161 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5221 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5281 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5341 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5401 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5461 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5521 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5581 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5641 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5701 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5761 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5941 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6001 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6061 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6121 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6181 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6241 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6301 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6361 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6421 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6481 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6541 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6601 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6661 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6721 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6781 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6841 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6901 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6961 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     7021 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7081 gct
//
