LOCUS       sequence-449150-        7669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of MpPHOT-Citrine in plant cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7669)
  AUTHORS   Yong LK, Keino I, Kanna Y, Noguchi M, Fujisawa M, Kodama Y
  TITLE     Functional comparison of phototropin from the liverworts Apopellia 
            endiviifolia and Marchantia polymorpha.
  JOURNAL   Photochem Photobiol. 2023 Oct 26. doi: 10.1111/php.13869.
  PUBMED    37882095
REFERENCE   2  (bases 1 to 7669)
  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: "Photochem 
            Photobiol. 2023 Oct 26. doi: 10.1111/php.13869."
COMMENT     Sequence Label: pGWT35S-MpPHOT-Citrine
FEATURES             Location/Qualifiers
     source          1..7669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2293..2295,2296..2298,2299..3009)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2338..2359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2599..2618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2946..2967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3014..3038)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      3083..3333
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3398..3416)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3399..3416)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3434..3450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3434..3450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3447..3469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3458..3474
                     /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(join(3482..3488,3489..3506,3507..3512))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3527..3548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3836..4424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3916..3935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4582..5397)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTSAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5305..5324
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5389..5408
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5582..5600
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5638..5660)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5760..5779
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5973..5995
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5987..6004
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5988..6004
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6014..6033
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6014..6032
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6109..6453
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     6396..6416
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    6473..6528
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    6572..6596
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 cttctgaatt acaggaaaga cggaactcct ttctggaacc ttctgacaat tgcacctatc
       61 aaggacgaca ctgggaaagt gctcaaatac attggaatgc aggtcgaggt cagcaaatac
      121 acagaaggaa acaagtccaa tgaagttcgg ccgaacggaa tgccggcttc acttatcaaa
      181 tacgacgctc ggcagcagga cagagccacg agttctgttc aggaactggt gggagctttg
      241 aaacatcctc accaatcacc gcaagataag ccctcggaag gaggcatgca atcgatgttt
      301 tcagttcctc cattgaaaga cgaactgcag ttgaaggtac cctcacgctt ttcaacgcag
      361 tcggttgtcg cactgccgct cgagggtagc caatcggcga aagcaagggc aaaaagtgag
      421 tcagtgagtc gactgaccga tgtcccagaa caacctggaa tggctgatgg cggccttcga
      481 ggggggcaga gccaccgcag atcgtcgggg ttgttgcaaa tgctcaggaa accgaaatca
      541 caacagccag aagctgatgt cgtcgacatc gatgacttgg accttgacga tgatgcccca
      601 cagaccatgg atgagaggcc agagagtatt gatgacagtg agcgagccaa ggaaattcgt
      661 aggggaatgg atttggccac aacgttagag cgaattgaga agaatttcgt gattacagat
      721 cctcgattgc cggataaccc cattattttc gcatccgacc acttcttaga gttaacggag
      781 tatactcgag aagaaattat cggaagaaat tgtcggtttc tgcaagggcc cgatacggac
      841 atggatgttg ttcggaaaat tagtgatgct atcaaacaac agcaaaacat cactgttcaa
      901 cttctgaatt acaccaaagg cggaaaaccg ttttggaatt tgtttcactt ggaagcaatg
      961 aaggacaaca agggtgagtt gcaatatttt atcggagtcc agctcgatgg aagtgagcac
     1021 atcgaaccca tcaggcgacg actctccgaa aagactgaag aggaaggcaa aaggattgtg
     1081 caagctacag cgaagaatgt tgacggagca gtgagggagc ttccagatgc gaatatgagc
     1141 atcgaggatt tgtgggcgaa tcactctcgt gttgtctttc ctaggcctca caaacgtcag
     1201 agttccacat ggcatgccat gcgaaagata ctcagttctg gagacaaatt gggtctcaac
     1261 cacttcaggc caatcaaacc tcttggttgt ggagataccg gcagtgtaca tctggtggag
     1321 ctgagaggga cgagtgagtt ctatgccatg aaagccatgg ataagaatgt tatggtgaat
     1381 cggaataagg ttcaccgtgc tcgggcagag agagaaattc tggagaaaat ggatcatccg
     1441 tttttgccca cgttatatgg ctccttccag actaggacac atgtgtgtct catcaccgat
     1501 ttttgtcctg gtggtgagct gttcttgctt ttggaaagac aaccccagaa gaagttccgt
     1561 gaggagtctg ctaggtttta tgcagccgag attgttcttg ccctcgaata tcttcactgc
     1621 atgggtgttg tatacaggga cctgaaacct gaaaacgttc tggtccaaaa agatgggcat
     1681 gtgcagctca ctgatttcga cctttccttc ttgactgcct ctcgtcctca gctgctgaaa
     1741 ccagtcgtac catctggccg aaagaataga cgagcaaggg agaacctacg accaatcctc
     1801 cttacagagc caatagcaaa gtctaactcc ttcgttggaa ctgaggagta tattgcgccg
     1861 gaaattatcc ttggtttggg acacaacagc tctgtagatt ggtgggctct aggtattctt
     1921 ctgtacgaga tgttatttgg acgaacaccc tttagaggac gcaaccggca aaggactttc
     1981 gcaaatgttc tgcataaaga tcttgccttc ccaagctcca tacctgtaag cttggctgcc
     2041 aaacaagtta ttcgtgggct gctacagaga gatcccaaga agaggttggg gtcagacaag
     2101 ggagcccacg acttaaaaca acatcaattc ttccggggca tcaattggcc tctcatccgc
     2161 tgcatgactc ctccaccgct agaaacacca atctaccaaa tcggcaaaga agcggactcc
     2221 aaagacctgg actgggagga gttggatccc agtgccgcct catttgatga atatggaggc
     2281 tcaggaggct ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2341 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2401 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2461 ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc
     2521 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2581 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2641 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2701 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2761 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2821 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2881 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     2941 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3001 ctgtacaagt gagacccagc tttcttgtac aaagtggttg gaggctcagg aggctcccca
     3061 tgggctagca ctagtatcta gaatcgttca aacatttggc aataaagttt cttaagattg
     3121 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat
     3181 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc
     3241 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa
     3301 ttatcgcgcg cggtgtcatc tatgttacta gatgcggccg caattgtgga tccgctctag
     3361 agtcgacctg caggcatgca agcttgcggc cgcgtattct atagtgtcac ctaaatagca
     3421 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3481 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4561 tctaaagtat atatgtttta attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     4621 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     4681 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     4741 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     4801 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aatagcttat gcatttcttt
     4861 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     4921 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     4981 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     5041 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc
     5101 agaggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     5161 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     5221 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     5281 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     5341 catgttggaa tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcataac
     5401 accccttgta ttactgttta tgtaagcaga cataagcatt tatcagggtt attgtctcat
     5461 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5521 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5581 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5641 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5701 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5761 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5821 cgtaaggaga aaatattact acaggcgcca ttcgccattc aggctgcgca actgttggga
     5881 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5941 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     6001 cagtgagcta gtgtaatacg actcactata gggcgcggcc gcagaattcg agctcggtac
     6061 ccgggatctc gaggccagat ctcccgggct cagaagaccg agggctattg agacttttca
     6121 acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttatc
     6181 gaaaggacag tagaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag
     6241 gctatcgttc aagatgcctc taccgacagt ggtcccaaag atggaccccc acccacgagg
     6301 aacatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat
     6361 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
     6421 atataaggaa gttcatttca tttggagagg acaggcttct tgagatcctt caacaattac
     6481 caacaacaac aaacaacaaa caacattaca attactattt acaattacag aattcgagct
     6541 cggtaccaag cttggaggct caggaggctc cacaagtttg tacaaaaaag caggcttcat
     6601 gatgccctcc acggattcat cgtctgcgaa gccttacaat ctgcgcaagc aaggggacac
     6661 gcccacaaaa cccgacgtgg gtcgagacgc tcgaggctct ctcgaagtat tcggcggggc
     6721 gccacaaacc tcagcattgc tgttctcgag gtcagctgcg gaagatgcct tggcagcggc
     6781 cggaattcga ccctcgcaaa ttctgagccc ctcgggctcc ggcaggcttc tgcccgcgct
     6841 cggcccgtta gggtcggacg gacaggcgct gccgctgtcg cagtccatgc tcaaggagcg
     6901 cgagaagcaa gttctgggca cagagcgcgg cgtgagaacg gactcgggac gcgccaggga
     6961 tcaagtggac aggtggctgg ctactgtcga cgagcgcgct ccgccttcca aaggctcggc
     7021 caaggagccg agcacggatt tctcgttgga cgacctggag ctgccatctt ccggcgctgc
     7081 tgctggccca ggagccagag gcgcagcagc cttggcctcg cccgaatggg ctggggccaa
     7141 aggcaaagct gtggtcgaga aggagccgag cgaagtggcc ggaggaggct tccaggcgcc
     7201 cgaaggtgtg tttcccctgg ggggcgacaa ggtggtgccc atggtgtcgg gcccgtcgga
     7261 cttgtcagag gaggtgcagg ccgagcgagc cgcgcaatgg ggagtgctga tgaagagcac
     7321 ggcagatctg gcctcgaccg gccgcgcgtc cgggtccaag gacgagcgga tttcggcctc
     7381 cggcatgtcc cgcagaactt cggaaggatc cgaaggcgag gttcctcgtg tctctagaga
     7441 tttgaaagat gctctcgctt ctttccagca gacctttgtg gtctcggatg ccacacgcac
     7501 ggacttcccc attctgtatg ccagtgcggg atttttcgcc atgacaggtt acactcccag
     7561 agaggtcatc ggaagaaact gtcgatttct ccagggacct gataccgacc aaagcgacat
     7621 agagagaatt cgaactgctc tcaaggaggg gaagagttat tgtggaaga
//
