LOCUS       sequence-114201-        7964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The rhodopsin-guanylyl cyclase of the aquatic fungus Blastocladiella
            emersonii enables fast optical control of cGMP signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7964)
  AUTHORS   Scheib U, Stehfest K, Gee CE, Korschen HG, Fudim R, Oertner TG, 
            Hegemann P
  TITLE     The rhodopsin-guanylyl cyclase of the aquatic fungus Blastocladiella
            emersonii enables fast optical control of cGMP signaling.
  JOURNAL   Sci Signal. 2015 Aug 11;8(389):rs8. doi: 10.1126/scisignal.aab0611.
  PUBMED    26268609
REFERENCE   2  (bases 1 to 7964)
  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"; journalName: "Sci 
            Signal."; date: "2015-08-11"; volume: "8(389):rs8. doi"; pages: " 
            10.1126/scisignal.aab0611"
COMMENT     Sequence Label: pAAV Syn hBE Rh Guanylyl Cyclase1 2A tDimer
FEATURES             Location/Qualifiers
     source          1..7964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..474
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2388..2441
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      2457..2466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2461..2470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2463..3140
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             3183..3857
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     misc_feature    3885..4473
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3938..3958)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4356..4367)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4476..4492)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4477..4493)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4553..5029
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4752..4771)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5069..5209
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5069..5198
                     /label=AAV2 ITR
     rep_origin      5284..5739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5371..5390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5581..5602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5756..5775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5875..5897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5935..5953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6021..6125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6126..6194,6195..6986)
                     /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     complement(6344..6363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7157..7745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7646..7665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7807..7947
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7818..7947
                     /label=AAV2 ITR
ORIGIN
        1 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
       61 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      121 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      181 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
      241 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
      301 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
      361 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
      421 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat
      481 tcgcaataat gtctatgaaa gataaggata ataacctgag aggagcttgt tctagctgta
      541 actgccctga atactgtttt agcccaactt ccaccctgtg cgacgattgc aagtgttccg
      601 tgaccaaaca cccaatcgtc gaacagcccc tgtcccgaaa cggatctttc cggagctccg
      661 gagcaagtct gctgccatca ccttctagtc ccaatgtgaa gattacctct acagtcggcc
      721 tgcggagtag aaaaagcgag tcccaggcaa acgtgcgcgg gtctatgatc tctaacagta
      781 attcaggaag ccgatccaac aatagtggag gagcaggagg agggtcagga ggctcaagct
      841 cctctaaggg gggcagcgcc ctggctaact accagtccgc tatgtctgag ctgtggtctt
      901 ggaatatgat gctgagtact ccttcactga aattcctgac cgtgcagttt accacatgga
      961 ttgtgctgac taccgtcggc gccatctaca ctctgttctt tcatgaaaga caggcttata
     1021 atagggggtg ggcagacatc tggtacggct atggggcttt cggatttggc ctggggctga
     1081 gcttcgcata tatgggcttc accggcgcca gaaacccaga gaagaaagct ctgagcctgt
     1141 gcctgctggg ggtgaatttc attagcttta tgtcctacat catcatcatg ctgcggctga
     1201 ctcccaccat cgaaggcacc atggccaacc cagtggagcc cgctagatat ctggaatgga
     1261 tcgccacttg ccctgtcctg atcctgctga ttagcgagat cacccagtac cctcacgacc
     1321 catataaagt gatcgtcaat gattacgccc tgtgcctggc aggattcgtg ggagcaatca
     1381 gcgcccagca gccatggggc gacctggcac atttcgtgag ctgcctgtgc ttttcctacg
     1441 tggtctattc tctgtggagt tgcttcacag gggctatcga tggagagact cagtgtaacg
     1501 tggaaaaaag cggcctgagg tggattcgct tttcaacaat cacaacttgg agcctgttcc
     1561 ccattacctg gttttcatat acaagcggcc tgatctcttt cactgtggca gaggccgggt
     1621 ttagtatgat tgacatcgga gccaaggtct tcctgaccct ggtgctggtc aacagcacag
     1681 tggaacaggc acagaatcag aaagtcgacg ccattaccgc tatcgcagag gaactggaga
     1741 accagattaa caattgtgat gccatcctgc agaagatgat gcccgagggc gtgctggaac
     1801 agctgaagaa tgggcaggct actgaggcaa aagagtacga atccgtgacc gtcttctttt
     1861 ctgatattac taactttacc gtgatcagtt caaggacatc cactaaggac atgatggcca
     1921 ccctgaacaa gctgtggctg gaatatgatg ccattgctaa gcggtgggga gtgtataaag
     1981 tcgagacaat cggcgacgct tatctgggag tgactggagc accagacgtg gtccctgatc
     2041 acgcagagcg agcttgcaac ttcgccgtgg atatcatcga aatgattaag agctttaaaa
     2101 ccatcacagg cgagtccatt aacatccgca ttggcctgaa tagcggacct gtgaccgcag
     2161 gagtcctggg cgacctgaac ccacactggt gtctggtggg agatacagtc aatactgcct
     2221 cccggatgga atctacaagt aaagctggcc acatccatat ttcagagagc acctaccatt
     2281 tcatcaagtc taaatttgtg acacagcctc tggatgtgat ggaggtgaaa ggcaaaggca
     2341 aaatgcagac ttattgggtg ctgggcagaa aatcggtacc cgcggccgag ggcagaggaa
     2401 gtcttctaac atgcggtgac gtggaggaga atcccggccc tgcaccggga tccgcggccg
     2461 ccatggtggc ctcctccgag gacgtcatca aagagttcat gcgcttcaag gtgcgcatgg
     2521 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     2581 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     2641 acatcctgtc cccccagttc cagtacggct ccaaggcgta cgtgaagcac cccgccgaca
     2701 tccccgacta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     2761 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     2821 tctacaaggt gaagttccgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     2881 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     2941 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     3001 ccatctacat ggccaagaag cccgtgcagc tgcccggcta ctactacgtg gacaccaagc
     3061 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     3121 gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc agctccggca
     3181 ccgcctcctc cgaggacgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct
     3241 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3301 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     3361 tgtcccccca gttccagtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg
     3421 actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3481 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca
     3541 aggtgaagtt ccgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca
     3601 tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga
     3661 tccaccaggc cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct
     3721 acatggccaa gaagcccgtg cagctgcccg gctactacta cgtggacacc aagctggaca
     3781 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc
     3841 accacctgtt cctgtaggtc gacttcgagc aaaagcttat cgataatcaa cctctggatt
     3901 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3961 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4021 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4081 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     4141 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4201 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4261 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     4321 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4381 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4441 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacgcga
     4501 tcgcggcgcg ccggcctctc tggcctcgac ctcgagtcta gagatgatct acgggtggca
     4561 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     4621 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     4681 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     4741 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     4801 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     4861 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     4921 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4981 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     5041 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     5101 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     5161 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     5221 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     5281 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5341 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5401 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5461 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5521 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5581 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     5641 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5701 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5761 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5821 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5881 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5941 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6001 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6061 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7801 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     7861 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     7921 tggccaactc catcactagg ggttcctgcg gccgcacgcg tcga
//
