LOCUS       sequence-193104-        7062 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  This plasmid is for expression of a rhodopsin/guanylyl cyclase 
            fusion protein mutant with lambda max = 390 nm..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7062)
  AUTHORS   Trieu MM, Devine EL, Lamarche LB, Ammerman AE, Greco JA, Birge RR, 
            Theobald DL, Oprian DD
  TITLE     Expression, purification, and spectral tuning of RhoGC, a 
            retinylidene/guanylyl cyclase fusion protein and optogenetics tool 
            from the aquatic fungus Blastocladiella emersonii.
  JOURNAL   J Biol Chem. 2017 Jun 23;292(25):10379-10389. doi: 
            10.1074/jbc.M117.789636. Epub 2017 May 4.
  PUBMED    28473465
REFERENCE   2  (bases 1 to 7062)
  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: 
            "10.1074/jbc.M117.789636"; journalName: "J Biol Chem"; date: 
            "2017-06-23- 23"; volume: "292"; issue: "25"; pages: "10379-10389"
COMMENT     Sequence Label: pMT3-RhoGC E254D
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             95..658
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    845..979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(882..901)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     936..955
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1652..1669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1823..2411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1903..1922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2582..3373,3374..3442))
                     /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     3205..3224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3443..3547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3615..3633
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3671..3693)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3793..3812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4006..4028
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4020..4037
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4021..4037
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4047..4376)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4061..4196
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4361..4381
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          4845..5074
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5057..5076
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gggggggggg ggacagctca gggctgcgat ttcgcgccaa acttgacggc aatcctagcg
       61 tgaaggctgg taggatttta tccccgctgc catcatggtt cgaccattga actgcatcgt
      121 cgccgtgtcc caaaatatgg ggattggcaa gaacggagac ctaccctggc ctccgctcag
      181 gaacgagttc aagtacttcc aaagaatgac cacaacctct tcagtggaag gtaaacagaa
      241 tctggtgatt atgggtagga aaacctggtt ctccattcct gagaagaatc gacctttaaa
      301 ggacagaatt aatatagttc tcagtagaga actcaaagaa ccaccacgag gagctcattt
      361 tcttgccaaa agtttggatg atgccttaag acttattgaa caaccggaat tggcaagtaa
      421 agtagacatg gtttggatag tcggaggcag ttctgtttac caggaagcca tgaatcaacc
      481 aggccacctc agactctttg tgacaaggat catgcaggaa tttgaaagtg acacgttttt
      541 cccagaaatt gatttgggga aatataaact tctcccagaa tacccaggcg tcctctctga
      601 ggtccaggag gaaaaaggca tcaagtataa gtttgaagtc tacgagaaga aagactaaca
      661 ggaagatgct ttcaagttct ctgctcccct cctaaagcta tgcattttta taagaccatg
      721 ggacttttgc tggctttaga tcataatcag ccataccaca tttgtagagg ttttacttgc
      781 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
      841 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      901 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
      961 atcttatcat gtctggatcc ccggccaacg gtctggtgac ccggctgcga gagctcggtg
     1021 tacctgagac gcgagtaagc ccttgagtca aagacgtagt cgttgcaagt ccgcaccagg
     1081 tactgatatc ccaccaaaaa gtgcggcggc ggctggcggt agaggggcca gcgtagggtg
     1141 gccggggctc cgggggcgag gtcttccaac ataaggcgat gatatccgta gatgtacctg
     1201 gacatccagg tgatgccggc ggcggtggtg gaggcgcgcg gaaagtcgcg gacgcggttc
     1261 cagatgttgc gcagcggcaa aaagtgctcc atggtcggga cgctctggcc ggtgaggcgt
     1321 gcgcagtcgt tgacgctcta gaccgtgcaa aaggagagcc tgtaagcggg cactcttccg
     1381 tggtctggtg gataaattcg caagggtatc atggcggacg accggggttc gaaccccgga
     1441 tccggccgtc cgccgtgatc catgcggtta ccgcccgcgt gtcgaaccca ggtgtgcgac
     1501 gtcagacaac gggggagcgc tccttttggc ttccttccag gcgcggcggc tgctgcgcta
     1561 gcttttttgg cgagctcgaa ttaattctgc attaatgaat cggccaacgc gcggggagag
     1621 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1681 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2341 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3541 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     3601 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     3661 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     3721 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     3781 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     3841 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     3901 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     3961 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     4021 gtaaaacgac ggccagtgcc aagctttttg caaaagccta ggcctccaaa aaagcctcct
     4081 cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa
     4141 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     4201 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     4261 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     4321 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     4381 cacaggatcc ggtcgcgcga atttcgagcg gtgttccgcg gtcctcctcg tatagaaact
     4441 cggaccactc tgagacgaag gctcgcgtcc aggccagcac gaaggaggct aagtgggagg
     4501 ggtagcggtc gttgtccact agggggtcca ctcgctccag ggtgtgaaga cacatgtcgc
     4561 cctcttcggc atcaaggaag gtgattggtt tataggtgta ggccacgtga ccgggtgttc
     4621 ctgaaggggg gctataaaag ggggtggggg cgcgttcgtc ctcactctct tccgcatcgc
     4681 tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca aactcttcgc ggtctttcca
     4741 gtactcttgg atcggaaacc cgtcggcctc cgaacggtac tccgccaccg agggacctga
     4801 gcgagtccgc atcgaccgga tcggaaaacc tctcgactgt tggggtgagt actccctctc
     4861 aaaagcgggc atgacttctg cgctaagatt gtcagtttcc aaaaacgagg aggatttgat
     4921 attcacctgg cccgcggtga tgcctttgag ggtggccgcg tccatctggt cagaaaagac
     4981 aatctttttg ttgtcaagct tgaggtgtgg caggcttgag atctggccat acacttgagt
     5041 gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcaa
     5101 gaattccacc atggccaaag acaaagacaa taatctgcgc ggcgcctgca gtagctgcaa
     5161 ctgtccggaa tattgcttta gcccgaccag taccctgtgc gacgattgta agtgcagcgt
     5221 gaccaaacat ccgattgttg agcagccgct gagtcgcaat ggtagctttc gcagtagtgg
     5281 tgcaagcctg ctgccgagcc cgagcagtcc gaatgtgaaa attaccagca ccgtgggtct
     5341 gcgcagtcgc aagagcgaga gccaggccaa tgtgcgcggc agcatgatta gcaacagcaa
     5401 tagcggcagc cgtagtaaca acagtggtgg cgccggtggt ggtagtggcg gtagcagtag
     5461 cagcaaaggc ggcagcgcat tagcaaatta tcagagcgca atgagcgaac tgtggagctg
     5521 gaacatgatg ctgagcaccc cgagtctgaa gtttctgacc gttcagttta ccacctggat
     5581 tgttctgacc accgttggcg ccatctacac cctgttcttc catgagcgtc aggcctacaa
     5641 ccgtggctgg gccgatattt ggtacggcta tggcgccttt ggcttcggcc tgggcctgag
     5701 ttttgcctat atgggcttta ccggcgcacg caaccctgaa aagaaggccc tgagcttatg
     5761 cctgctgggt gttaatttca ttagctttat gagttatatc attatcatgt tacgcctgac
     5821 cccgaccatc gaaggtacta tggccaatcc ggtggaaccg gcacgctatc tggactggat
     5881 cgccacctgc ccggtgctga ttctgctgat tagtgagatc acccagtatc cgcacgaccc
     5941 gtataaagtg atcgtgaacg attacgcact gtgcctggca ggtttcgtgg gtgcaatcag
     6001 cgcacagcag ccgtggggcg atctggccca ttttgtgagc tgcctgtgct tcagctacgt
     6061 ggtgtatagc ctgtggagtt gctttacagg tgccatcgat ggcgaaaccc agtgcaatgt
     6121 ggaaaaaagc ggcctgcgct ggattcgttt tagcaccatc accacctgga gcctgtttcc
     6181 tatcacctgg ttcagctaca ccagtggtct gattagtttt accgtggcag aggcaggctt
     6241 cagcatgatt gacattggcg ccaaagtgtt tctgacactg gtgctggtga atagcaccgt
     6301 ggagcaagcc cagaatcaaa aggtggacgc cattaccgcc attgccgagg agctggagaa
     6361 tcagattaat aactgtgatg caatcctgca aaaaatgatg ccggaaggcg tgttagagca
     6421 gctgaagaat ggtcaggcca ccgaagccaa agaatacgaa agcgttaccg tgttcttcag
     6481 tgacatcacc aactttaccg tgattagcag ccgtaccagc accaaggata tgatggccac
     6541 cctgaacaag ttatggctgg agtatgatgc aatcgccaag cgctggggtg tgtataaggt
     6601 ggagacaatc ggcgacgcat atctgggtgt taccggtgca ccggatgtgg ttccggatca
     6661 tgcagagcgc gcctgcaatt ttgccgtgga catcattgag atgatcaaaa gctttaagac
     6721 aattaccggc gaaagcatta acattcgtat tggcttaaat agcggcccgg tgaccgcagg
     6781 cgtgttaggt gatctgaacc cgcattggtg cctggttggt gacaccgtta ataccgccag
     6841 tcgtatggag agcaccagta aagccggcca cattcatatt agtgagagca cctatcattt
     6901 tattaaaagc aaattcgtga cccagccgct ggacgtgatg gaagtgaagg gcaaaggcaa
     6961 gatgcagaca tactgggttc tgggtcgcaa aatgcatggc agcggcagca cagagaccag
     7021 ccaagtggcg cctgcctaag cggccgcaaa ttccgggggc tg
//
