LOCUS       sequence-193105-        7053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is for expression of a rhodopsin/guanylyl cyclase 
            fusion protein mutant with adenylyl cyclase activity. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7053)
  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 7053)
  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-RhoAC
FEATURES             Location/Qualifiers
     source          1..7053
                     /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             2749..3312
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    3499..3633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3536..3555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3590..3609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4306..4323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4477..5065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4557..4576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5236..6027,6028..6096))
                     /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     5859..5878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6097..6201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6269..6287
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6325..6347)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6447..6466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6660..6682
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6674..6691
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6675..6691
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6701..7030)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6715..6850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7015..7035
                     /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 tgacattggc
     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 tgaagacaat cggcgacgca
     2221 tatctgggtg ttaccggtgc accggatgtg gttccggatc atgcagagcg cgcctgcaat
     2281 tttgccgtgg acatcattga gatgatcaaa agctttaaga caattaccgg cgaaagcatt
     2341 aacattcgta ttggcttaaa tagcggcccg gtgaccgcag gcgtgttagg tgatctgaac
     2401 ccgcattggg acctggttgg 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 ggggggacag ctcagggctg cgatttcgcg ccaaacttga
     2701 cggcaatcct agcgtgaagg ctggtaggat tttatccccg ctgccatcat ggttcgacca
     2761 ttgaactgca tcgtcgccgt gtcccaaaat atggggattg gcaagaacgg agacctaccc
     2821 tggcctccgc tcaggaacga gttcaagtac ttccaaagaa tgaccacaac ctcttcagtg
     2881 gaaggtaaac agaatctggt gattatgggt aggaaaacct ggttctccat tcctgagaag
     2941 aatcgacctt taaaggacag aattaatata gttctcagta gagaactcaa agaaccacca
     3001 cgaggagctc attttcttgc caaaagtttg gatgatgcct taagacttat tgaacaaccg
     3061 gaattggcaa gtaaagtaga catggtttgg atagtcggag gcagttctgt ttaccaggaa
     3121 gccatgaatc aaccaggcca cctcagactc tttgtgacaa ggatcatgca ggaatttgaa
     3181 agtgacacgt ttttcccaga aattgatttg gggaaatata aacttctccc agaataccca
     3241 ggcgtcctct ctgaggtcca ggaggaaaaa ggcatcaagt ataagtttga agtctacgag
     3301 aagaaagact aacaggaaga tgctttcaag ttctctgctc ccctcctaaa gctatgcatt
     3361 tttataagac catgggactt ttgctggctt tagatcataa tcagccatac cacatttgta
     3421 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3481 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3541 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3601 aaactcatca atgtatctta tcatgtctgg atccccggcc aacggtctgg tgacccggct
     3661 gcgagagctc ggtgtacctg agacgcgagt aagcccttga gtcaaagacg tagtcgttgc
     3721 aagtccgcac caggtactga tatcccacca aaaagtgcgg cggcggctgg cggtagaggg
     3781 gccagcgtag ggtggccggg gctccggggg cgaggtcttc caacataagg cgatgatatc
     3841 cgtagatgta cctggacatc caggtgatgc cggcggcggt ggtggaggcg cgcggaaagt
     3901 cgcggacgcg gttccagatg ttgcgcagcg gcaaaaagtg ctccatggtc gggacgctct
     3961 ggccggtgag gcgtgcgcag tcgttgacgc tctagaccgt gcaaaaggag agcctgtaag
     4021 cgggcactct tccgtggtct ggtggataaa ttcgcaaggg tatcatggcg gacgaccggg
     4081 gttcgaaccc cggatccggc cgtccgccgt gatccatgcg gttaccgccc gcgtgtcgaa
     4141 cccaggtgtg cgacgtcaga caacggggga gcgctccttt tggcttcctt ccaggcgcgg
     4201 cggctgctgc gctagctttt ttggcgagct cgaattaatt ctgcattaat gaatcggcca
     4261 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4321 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4381 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5221 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5281 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5341 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5401 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5461 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5521 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5581 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5641 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5701 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5761 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5821 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5881 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5941 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6001 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6061 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6241 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6301 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6361 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6421 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6481 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6541 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6601 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6661 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt tttgcaaaag cctaggcctc
     6721 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     6781 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     6841 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     6901 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     6961 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     7021 actgacacac attccacagg atccggtcgc gcg
//
