LOCUS       Exported                6788 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  1-621 amino acids of Arabidopsis PHYB (Gene ID: 816394), tagged with
            mCherry fluorescence protein and the CAAX membrane moiety. Mammalian
            codon optimised..
ACCESSION   .
VERSION     .
KEYWORDS    pCS2+-N-PAS2-GAF-PHYB-MCherry-CAAX
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6788)
  AUTHORS   Buckley CE, Moore RE, Reade A, Goldberg AR, Weiner OD, Clarke JD
  TITLE     Reversible Optogenetic Control of Subcellular Protein Localization 
            in a Live Vertebrate Embryo.
  JOURNAL   Dev Cell. 2016 Jan 11;36(1):117-26. doi: 
            10.1016/j.devcel.2015.12.011.
  PUBMED    26766447
REFERENCE   2  (bases 1 to 6788)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6788
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      109..118
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(2818..2835)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2840..2974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2864..2883)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2918..2937
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3087..3107)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3087..3105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3126..3142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3126..3142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3139..3161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3150..3166
                     /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(3174..3204)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3219..3240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3357..3374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3528..4116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3608..3627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4287..5147)
                     /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     4910..4929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5148..5252)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5278..5733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5415..5436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5627..5646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5811..6788
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggcgcgtgc caccatggta
      121 tcaggtgttg gaggaagtgg tggaggtaga ggtggaggaa gaggtggaga agaagagccc
      181 tcaagttcac acactcccaa caatagacga gggggtgaac aagcgcagtc ttctggtacg
      241 aagtccctcc ggcctcgctc taatacagaa tccatgtcca aagccatcca gcagtataca
      301 gtggatgcca ggctccacgc agtttttgaa cagagcgggg aaagtgggaa atctttcgat
      361 tatagccaga gtctgaagac cacaacttac ggcagcagtg ttccagagca acagattacg
      421 gcttacctta gccggataca gcgcggaggc tacatacagc cttttgggtg tatgatcgcg
      481 gttgacgaat ctagcttccg cataatcgga tattctgaaa acgctcggga gatgctgggt
      541 atcatgccac agagcgttcc tacactggaa aagcctgaaa tcctcgcgat gggcaccgat
      601 gtgcgcagcc tcttcacgtc atcttcaagt atcctcctgg agcgcgcttt tgttgcccgg
      661 gaaatcacac ttctgaatcc tgtctggatt catagcaaga atacagggaa acccttttac
      721 gctatcctgc acagaatcga tgtcggggtt gtaatagacc ttgagccagc tagaacagaa
      781 gatccggcct tgagcatcgc tggcgcagtg caaagccaga agctggcagt gagggcaata
      841 tctcaactgc aagccctgcc aggcggcgat ataaagctgc tgtgtgatac cgtagtggag
      901 agcgttcgcg atctgaccgg ttatgacaga gtgatggtct acaaatttca tgaagatgag
      961 catggggagg tagtggcaga atcaaaaaga gacgacttgg aaccctatat cggactccac
     1021 tatcctgcta ctgatattcc ccaggcaagt cggtttttgt ttaagcagaa tagggtcagg
     1081 atgattgtgg actgtaatgc tactcccgtg ttggtagtcc aagacgatcg actgacccaa
     1141 tctatgtgcc tggtggggag tacactgcgc gcaccacacg ggtgccattc ccagtatatg
     1201 gcgaacatgg ggagcatcgc ctcactggca atggctgtaa tcatcaacgg taacgaggat
     1261 gatggatcta atgtcgccag tggcagatcc tcaatgcgcc tctgggggct ggtcgtctgc
     1321 catcatacaa gcagccggtg tattcccttt ccgctgaggt acgcttgtga gtttcttatg
     1381 caagcatttg ggcttcagct taatatggaa ctccagcttg cccttcaaat gagtgagaag
     1441 cgggttctcc gcactcagac ccttctctgc gatatgcttt tgcgcgattc ccccgctggc
     1501 atcgtgaccc agagcccttc aataatggac cttgtaaagt gcgatggtgc tgcattcctg
     1561 tatcatggga aatactatcc actgggagtg gccccaagcg aagtacagat aaaggacgtg
     1621 gtcgagtggc tgctcgccaa tcacgccgac agtacagggc tgagcaccga tagtctgggg
     1681 gacgccggat atccaggtgc cgccgcactt ggcgatgcag tgtgtggcat ggcagtcgcc
     1741 tatattacga agcgcgattt tctcttttgg tttcgatcac acacagcaaa agagataaag
     1801 tggggaggcg ccaagcacca tccagaggat aaggatgacg ggcaaagaat gcatccaaga
     1861 agttctttcc aagccttcct cgaagtggtt aagtcacgat cacagccttg ggaaaccgcc
     1921 gaaatggacg ctatccattc tctccagctg atcctgcggg acagtttcaa ggagagcgct
     1981 agcgattccg caggctctgc tggctcagca ggcctgcagg tgtctaaagg cgaggaggat
     2041 aacatggcta taatcaagga atttatgagg ttcaaggtgc atatggaggg atctgtgaac
     2101 ggtcacgagt tcgaaatcga gggcgaaggc gaggggcgac cctatgaagg gacacagacg
     2161 gccaaactga aggtgaccaa gggtggcccc ctgcctttcg cctgggacat cctgtcaccc
     2221 cagtttatgt acggctctaa ggcttacgtt aagcaccctg ccgatattcc ggactacttg
     2281 aaactgagct tccctgaagg gttcaagtgg gaacgggtaa tgaattttga ggatggcggt
     2341 gtagtcacag ttacccagga cagctctctt caggacggag aatttatcta taaggttaaa
     2401 cttcggggca ctaacttccc atcagacggc cccgtgatgc agaagaaaac tatgggctgg
     2461 gaagccagct cagaacgcat gtatcccgag gacggagcac tgaagggaga aatcaagcag
     2521 cgactcaagc tgaaggatgg gggacattat gatgctgagg tcaaaaccac ctataaggcc
     2581 aagaaacccg ttcagcttcc tggggcatac aacgtgaata tcaaactgga tattacatcc
     2641 cataacgaag actataccat cgtggaacag tacgagcggg ccgagggcag acatagcaca
     2701 gggggcatgg atgaattgta caagctcgag aaaatgtcca aggatggtaa gaaaaagaag
     2761 aagaagtcaa aaaccaagtg tgttatcatg taagcggccc ctctcgagcc tctagaacta
     2821 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     2881 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2941 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3001 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     3061 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     3121 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3181 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     5281 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc atctcatttt
     5341 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5401 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5461 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5521 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5581 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5641 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5701 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     5761 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc gccaattcaa
     5821 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa
     5881 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa
     5941 tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt gccaactggg
     6001 gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc
     6061 gccatattga attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca
     6121 atatataggc aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac
     6181 tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata
     6241 gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc
     6301 cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg
     6361 cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc
     6421 ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct
     6481 cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc
     6541 acttggcagt acatcaatat ctattaatag taacttggca agtacattac tattggaagt
     6601 acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg
     6661 gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc
     6721 cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt
     6781 tagggaac
//
