LOCUS       sequence-192913-        7058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is for expression of a rhodopsin/guanylyl cyclase 
            fusion protein mutant with lambda max = 533 nm..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7058)
  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 7058)
  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 D380E
FEATURES             Location/Qualifiers
     source          1..7058
                     /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             2754..3317
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    3504..3638
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3541..3560)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3595..3614
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4311..4328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4482..5070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4562..4581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5241..6032,6033..6101))
                     /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     5864..5883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6102..6206)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6274..6292
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6330..6352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6452..6471
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6665..6687
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6679..6696
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6680..6696
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6706..7035)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6720..6855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7020..7040
                     /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 tgagattggc
     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 gggggggggg gacagctcag ggctgcgatt tcgcgccaaa
     2701 cttgacggca atcctagcgt gaaggctggt aggattttat ccccgctgcc atcatggttc
     2761 gaccattgaa ctgcatcgtc gccgtgtccc aaaatatggg gattggcaag aacggagacc
     2821 taccctggcc tccgctcagg aacgagttca agtacttcca aagaatgacc acaacctctt
     2881 cagtggaagg taaacagaat ctggtgatta tgggtaggaa aacctggttc tccattcctg
     2941 agaagaatcg acctttaaag gacagaatta atatagttct cagtagagaa ctcaaagaac
     3001 caccacgagg agctcatttt cttgccaaaa gtttggatga tgccttaaga cttattgaac
     3061 aaccggaatt ggcaagtaaa gtagacatgg tttggatagt cggaggcagt tctgtttacc
     3121 aggaagccat gaatcaacca ggccacctca gactctttgt gacaaggatc atgcaggaat
     3181 ttgaaagtga cacgtttttc ccagaaattg atttggggaa atataaactt ctcccagaat
     3241 acccaggcgt cctctctgag gtccaggagg aaaaaggcat caagtataag tttgaagtct
     3301 acgagaagaa agactaacag gaagatgctt tcaagttctc tgctcccctc ctaaagctat
     3361 gcatttttat aagaccatgg gacttttgct ggctttagat cataatcagc cataccacat
     3421 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3481 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3541 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3601 tgtccaaact catcaatgta tcttatcatg tctggatccc cggccaacgg tctggtgacc
     3661 cggctgcgag agctcggtgt acctgagacg cgagtaagcc cttgagtcaa agacgtagtc
     3721 gttgcaagtc cgcaccaggt actgatatcc caccaaaaag tgcggcggcg gctggcggta
     3781 gaggggccag cgtagggtgg ccggggctcc gggggcgagg tcttccaaca taaggcgatg
     3841 atatccgtag atgtacctgg acatccaggt gatgccggcg gcggtggtgg aggcgcgcgg
     3901 aaagtcgcgg acgcggttcc agatgttgcg cagcggcaaa aagtgctcca tggtcgggac
     3961 gctctggccg gtgaggcgtg cgcagtcgtt gacgctctag accgtgcaaa aggagagcct
     4021 gtaagcgggc actcttccgt ggtctggtgg ataaattcgc aagggtatca tggcggacga
     4081 ccggggttcg aaccccggat ccggccgtcc gccgtgatcc atgcggttac cgcccgcgtg
     4141 tcgaacccag gtgtgcgacg tcagacaacg ggggagcgct ccttttggct tccttccagg
     4201 cgcggcggct gctgcgctag cttttttggc gagctcgaat taattctgca ttaatgaatc
     4261 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4321 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4381 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4441 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4621 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4801 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4921 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5041 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5101 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5161 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5221 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5281 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5341 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5401 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5461 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5521 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5581 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5641 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5701 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5761 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5821 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5881 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5941 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6001 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6061 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6121 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6181 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6241 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6301 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6361 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6421 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6481 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6541 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6601 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6661 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctttttgc aaaagcctag
     6721 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     6781 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6841 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     6901 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     6961 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     7021 ccctaactga cacacattcc acaggatccg gtcgcgcg
//
