LOCUS       sequence-193102-        7052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is for expression of a rhodopsin/guanylyl cyclase 
            fusion protein mutant with lambda max = 506 nm..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7052)
  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 7052)
  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 D380N
FEATURES             Location/Qualifiers
     source          1..7052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          446..675
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     658..677
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2748..3311
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    3498..3632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3535..3554)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3589..3608
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4305..4322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4476..5064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4556..4575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5235..6026,6027..6095))
                     /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     5858..5877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6096..6200)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6268..6286
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6324..6346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6446..6465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6659..6681
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6673..6690
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6674..6690
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6700..7029)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6714..6849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7014..7034
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aatttcgagc ggtgttccgc ggtcctcctc gtatagaaac tcggaccact ctgagacgaa
       61 ggctcgcgtc caggccagca cgaaggaggc taagtgggag gggtagcggt cgttgtccac
      121 tagggggtcc actcgctcca gggtgtgaag acacatgtcg ccctcttcgg catcaaggaa
      181 ggtgattggt ttataggtgt aggccacgtg accgggtgtt cctgaagggg ggctataaaa
      241 gggggtgggg gcgcgttcgt cctcactctc ttccgcatcg ctgtctgcga gggccagctg
      301 ttgggctcgc ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac
      361 ccgtcggcct ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg
      421 atcggaaaac ctctcgactg ttggggtgag tactccctct caaaagcggg catgacttct
      481 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
      541 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
      601 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
      661 cctttctctc cacaggtgtc cactcccagg tccaactgca agaattccac catggccaaa
      721 gacaaagaca ataatctgcg cggcgcctgc agtagctgca actgtccgga atattgcttt
      781 agcccgacca gtaccctgtg cgacgattgt aagtgcagcg tgaccaaaca tccgattgtt
      841 gagcagccgc tgagtcgcaa tggtagcttt cgcagtagtg gtgcaagcct gctgccgagc
      901 ccgagcagtc cgaatgtgaa aattaccagc accgtgggtc tgcgcagtcg caagagcgag
      961 agccaggcca atgtgcgcgg cagcatgatt agcaacagca atagcggcag ccgtagtaac
     1021 aacagtggtg gcgccggtgg tggtagtggc ggtagcagta gcagcaaagg cggcagcgca
     1081 ttagcaaatt atcagagcgc aatgagcgaa ctgtggagct ggaacatgat gctgagcacc
     1141 ccgagtctga agtttctgac cgttcagttt accacctgga ttgttctgac caccgttggc
     1201 gccatctaca ccctgttctt ccatgagcgt caggcctaca accgtggctg ggccgatatt
     1261 tggtacggct atggcgcctt tggcttcggc ctgggcctga gttttgccta tatgggcttt
     1321 accggcgcac gcaaccctga aaagaaggcc ctgagcttat gcctgctggg tgttaatttc
     1381 attagcttta tgagttatat cattatcatg ttacgcctga ccccgaccat cgaaggtact
     1441 atggccaatc cggtggaacc ggcacgctat ctggaatgga tcgccacctg cccggtgctg
     1501 attctgctga ttagtgagat cacccagtat ccgcacgacc cgtataaagt gatcgtgaac
     1561 gattacgcac tgtgcctggc aggtttcgtg ggtgcaatca gcgcacagca gccgtggggc
     1621 gatctggccc attttgtgag ctgcctgtgc ttcagctacg tggtgtatag cctgtggagt
     1681 tgctttacag gtgccatcga tggcgaaacc cagtgcaatg tggaaaaaag cggcctgcgc
     1741 tggattcgtt ttagcaccat caccacctgg agcctgtttc ctatcacctg gttcagctac
     1801 accagtggtc tgattagttt taccgtggca gaggcaggct tcagcatgat taacattggc
     1861 gccaaagtgt ttctgacact ggtgctggtg aatagcaccg tggagcaagc ccagaatcaa
     1921 aaggtggacg ccattaccgc cattgccgag gagctggaga atcagattaa taactgtgat
     1981 gcaatcctgc aaaaaatgat gccggaaggc gtgttagagc agctgaagaa tggtcaggcc
     2041 accgaagcca aagaatacga aagcgttacc gtgttcttca gtgacatcac caactttacc
     2101 gtgattagca gccgtaccag caccaaggat atgatggcca ccctgaacaa gttatggctg
     2161 gagtatgatg caatcgccaa gcgctggggt gtgtataagg tggagacaat cggcgacgca
     2221 tatctgggtg ttaccggtgc accggatgtg gttccggatc atgcagagcg cgcctgcaat
     2281 tttgccgtgg acatcattga gatgatcaaa agctttaaga caattaccgg cgaaagcatt
     2341 aacattcgta ttggcttaaa tagcggcccg gtgaccgcag gcgtgttagg tgatctgaac
     2401 ccgcattggt gcctggttgg tgacaccgtt aataccgcca gtcgtatgga gagcaccagt
     2461 aaagccggcc acattcatat tagtgagagc acctatcatt ttattaaaag caaattcgtg
     2521 acccagccgc tggacgtgat ggaagtgaag ggcaaaggca agatgcagac atactgggtt
     2581 ctgggtcgca aaatgcatgg cagcggcagc acagagacca gccaagtggc gcctgcctaa
     2641 gcggccgcaa attccggggg gggggacagc tcagggctgc gatttcgcgc caaacttgac
     2701 ggcaatccta gcgtgaaggc tggtaggatt ttatccccgc tgccatcatg gttcgaccat
     2761 tgaactgcat cgtcgccgtg tcccaaaata tggggattgg caagaacgga gacctaccct
     2821 ggcctccgct caggaacgag ttcaagtact tccaaagaat gaccacaacc tcttcagtgg
     2881 aaggtaaaca gaatctggtg attatgggta ggaaaacctg gttctccatt cctgagaaga
     2941 atcgaccttt aaaggacaga attaatatag ttctcagtag agaactcaaa gaaccaccac
     3001 gaggagctca ttttcttgcc aaaagtttgg atgatgcctt aagacttatt gaacaaccgg
     3061 aattggcaag taaagtagac atggtttgga tagtcggagg cagttctgtt taccaggaag
     3121 ccatgaatca accaggccac ctcagactct ttgtgacaag gatcatgcag gaatttgaaa
     3181 gtgacacgtt tttcccagaa attgatttgg ggaaatataa acttctccca gaatacccag
     3241 gcgtcctctc tgaggtccag gaggaaaaag gcatcaagta taagtttgaa gtctacgaga
     3301 agaaagacta acaggaagat gctttcaagt tctctgctcc cctcctaaag ctatgcattt
     3361 ttataagacc atgggacttt tgctggcttt agatcataat cagccatacc acatttgtag
     3421 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3481 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3541 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3601 aactcatcaa tgtatcttat catgtctgga tccccggcca acggtctggt gacccggctg
     3661 cgagagctcg gtgtacctga gacgcgagta agcccttgag tcaaagacgt agtcgttgca
     3721 agtccgcacc aggtactgat atcccaccaa aaagtgcggc ggcggctggc ggtagagggg
     3781 ccagcgtagg gtggccgggg ctccgggggc gaggtcttcc aacataaggc gatgatatcc
     3841 gtagatgtac ctggacatcc aggtgatgcc ggcggcggtg gtggaggcgc gcggaaagtc
     3901 gcggacgcgg ttccagatgt tgcgcagcgg caaaaagtgc tccatggtcg ggacgctctg
     3961 gccggtgagg cgtgcgcagt cgttgacgct ctagaccgtg caaaaggaga gcctgtaagc
     4021 gggcactctt ccgtggtctg gtggataaat tcgcaagggt atcatggcgg acgaccgggg
     4081 ttcgaacccc ggatccggcc gtccgccgtg atccatgcgg ttaccgcccg cgtgtcgaac
     4141 ccaggtgtgc gacgtcagac aacgggggag cgctcctttt ggcttccttc caggcgcggc
     4201 ggctgctgcg ctagcttttt tggcgagctc gaattaattc tgcattaatg aatcggccaa
     4261 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4321 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5221 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5281 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5341 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5401 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5461 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5521 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5581 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5641 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5701 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5761 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5821 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5881 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5941 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6001 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6061 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6121 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6181 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6241 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6301 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     6361 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     6421 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     6481 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     6541 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6601 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     6661 cagtcacgac gttgtaaaac gacggccagt gccaagcttt ttgcaaaagc ctaggcctcc
     6721 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     6781 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     6841 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     6901 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     6961 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     7021 ctgacacaca ttccacagga tccggtcgcg cg
//
