LOCUS       Exported                4923 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  The rhodopsin-guanylyl cyclase of the aquatic fungus Blastocladiella
            emersonii enables fast optical control of cGMP signaling.
ACCESSION   .
VERSION     .
KEYWORDS    RhGC(BE)_pGEM
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4923)
  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 4923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4923
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     33..55
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     47..64
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     48..64
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     71..90
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        71..89
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           121..163
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           2089..2214
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2335..2353)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2336..2353)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2371..2387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2371..2387)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2384..2406)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2395..2411
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2419..2449)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2464..2485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2602..2619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2773..3361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2853..2872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3532..4392)
                     /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     4155..4174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4393..4497)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4565..4583
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4621..4643)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4743..4762
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
       61 cagtgaattg taatacgact cactataggg cgaattaatt cgagctcggt acccagcttg
      121 cttgttcttt ttgcagaagc tcagaataaa cgctcaactt tggcagatca attccccggg
      181 gatccatgaa agataaggat aataacctga gaggagcttg ttctagctgt aactgccctg
      241 aatactgttt tagcccaact tccaccctgt gcgacgattg caagtgttcc gtgaccaaac
      301 acccaatcgt cgaacagccc ctgtcccgaa acggatcttt ccggagctcc ggagcaagtc
      361 tgctgccatc accttctagt cccaatgtga agattacctc tacagtcggc ctgcggagta
      421 gaaaaagcga gtcccaggca aacgtgcgcg ggtctatgat ctctaacagt aattcaggaa
      481 gccgatccaa caatagtgga ggagcaggag gagggtcagg aggctcaagc tcctctaagg
      541 ggggcagcgc cctggctaac taccagtccg ctatgtctga gctgtggtct tggaatatga
      601 tgctgagtac tccttcactg aaattcctga ccgtgcagtt taccacatgg attgtgctga
      661 ctaccgtcgg cgccatctac actctgttct ttcatgaaag acaggcttat aatagggggt
      721 gggcagacat ctggtacggc tatggggctt tcggatttgg cctggggctg agcttcgcat
      781 atatgggctt caccggcgcc agaaacccag agaagaaagc tctgagcctg tgcctgctgg
      841 gggtgaattt cattagcttt atgtcctaca tcatcatcat gctgcggctg actcccacca
      901 tcgaaggcac catggccaac ccagtggagc ccgctagata tctggaatgg atcgccactt
      961 gccctgtcct gatcctgctg attagcgaga tcacccagta ccctcacgac ccatataaag
     1021 tgatcgtcaa tgattacgcc ctgtgcctgg caggattcgt gggagcaatc agcgcccagc
     1081 agccatgggg cgacctggca catttcgtga gctgcctgtg cttttcctac gtggtctatt
     1141 ctctgtggag ttgcttcaca ggggctatcg atggagagac tcagtgtaac gtggaaaaaa
     1201 gcggcctgag gtggattcgc ttttcaacaa tcacaacttg gagcctgttc cccattacct
     1261 ggttttcata tacaagcggc ctgatctctt tcactgtggc agaggccggg tttagtatga
     1321 ttgacatcgg agccaaggtc ttcctgaccc tggtgctggt caacagcaca gtggaacagg
     1381 cacagaatca gaaagtcgac gccattaccg ctatcgcaga ggaactggag aaccagatta
     1441 acaattgtga tgccatcctg cagaagatga tgcccgaggg cgtgctggaa cagctgaaga
     1501 atgggcaggc tactgaggca aaagagtacg aatccgtgac cgtcttcttt tctgatatta
     1561 ctaactttac cgtgatcagt tcaaggacat ccactaagga catgatggcc accctgaaca
     1621 agctgtggct ggaatatgat gccattgcta agcggtgggg agtgtataaa gtcgagacaa
     1681 tcggcgacgc ttatctggga gtgactggag caccagacgt ggtccctgat cacgcagagc
     1741 gagcttgcaa cttcgccgtg gatatcatcg aaatgattaa gagctttaaa accatcacag
     1801 gcgagtccat taacatccgc attggcctga atagcggacc tgtgaccgca ggagtcctgg
     1861 gcgacctgaa cccacactgg tgtctggtgg gagatacagt caatactgcc tcccggatgg
     1921 aatctacaag taaagctggc cacatccata tttcagagag cacctaccat ttcatcaagt
     1981 ctaaatttgt gacacagcct ctggatgtga tggaggtgaa aggcaaaggc aaaatgcaga
     2041 cttattgggt gctgggcaga aaatagaagc ttgatctggt taccactaaa ccagcctcaa
     2101 gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt
     2161 cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattcta
     2221 aaaaaaaaaa aaaaaaaaaa aacccccccc cccccccccc cccccccccc ccctgcaggc
     2281 atgcgtttaa acatttaaat tacgtattaa ttaagatatc gctagcttga gtattctata
     2341 gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2401 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2461 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2521 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2581 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2641 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2701 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2761 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2821 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2881 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2941 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3001 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3061 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3121 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3181 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3241 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3301 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3361 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3421 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3481 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3841 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4141 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4441 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4501 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4561 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4621 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4681 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4741 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4801 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     4861 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     4921 tgc
//
