LOCUS       sequence-449147-        7573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of AePHOT-Citrine in plant cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7573)
  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 7573)
  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-AePHOT-Citrine
FEATURES             Location/Qualifiers
     source          1..7573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1422..1433)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1640..1664)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1708..1763
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     promoter        complement(1783..2127)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1820..1840)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     complement(2203..2222)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2204..2222)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2232..2249)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2232..2248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2241..2263)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2457..2476)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2576..2598
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2636..2654)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2828..2847)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2839..3654
                     /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     complement(2912..2931)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3812..4400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4301..4320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4554..4571
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4688..4709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4724..4729,4730..4747,4748..4754)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4762..4778
                     /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)."
     primer_bind     4767..4789
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4786..4802
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4786..4802
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4820..4838
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4820..4837
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(4903..5153)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    5198..5222
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(5227..5937,5938..5940,5941..5943))
                     /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(5269..5290)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5618..5637
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5877..5898
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 atgcctcgac gaatctccgt agcccgctct gaattatcca aactttctct gtcctccaga
       61 ttcggtacag aatcatctag atcaagatca tctacttctg gtggaggtgg tgcaggcttt
      121 gctggtttct taatcaactg catcagtccg gaaggccgac gagtgctttg gctcttccca
      181 gcatttcccc ccactgaacg tgggtctgct tcatctggag ttctgttcac tgattcactt
      241 tttgatctaa atttcggctg attatagtcc tcaggaatct tccccattgc agaagattga
      301 gttgaatagc gggaaggtaa cttcagctgt agctcatctt tcaagggtgg aaatgaaaac
      361 agtggtggag caccgccacc agtgcttttt cgtggaggtg atgcttggtg tccattacca
      421 tttggatgtt tgagagcacc gaccagttct tgaacagaac ttgccgccct gtcttgctgc
      481 cgagcatcat attttatcag ggatgctggc attccattgg gtcggacatc gtttgcccta
      541 tttccttcag tatacttgct aacttccacc tgcattccaa tgtatttcag aatcttccca
      601 ttgtcgtctt taatgggtgc tattgtaagg agattccaga aaggggttcc attctttctg
      661 taatttagaa gtctcccaca aaaagtcttc ccatccttca gtgcagatcg aatcctctca
      721 atatcttctt gatcagtttc cggtccctgg agaaaccgac agttcctccc gatgacctct
      781 ttcggtgcat atcctgtcat gtcgaagaac cccgcactgg catagaggat tgggtagtca
      841 ggacgggtag catcagacac tacaaaggtt tgctggaacg aagcaagggc atccttgaga
      901 tcccgagaaa tgcgagggac agtttgcgaa tcactccctc catcagatgc ttccgaagtc
      961 ctccctgaca atcccgatat ccgatcgtct cgggagcctg aggcacgccc agtcaaatct
     1021 gctgctacta ctgctttcaa tgctgctggc cattgcaatc ctctctctga acttgcctca
     1081 tcggaaggat cgggtattac tgattgcacg agtgctcctg ctccgttccc caattcagaa
     1141 tccttcccga cacgtttgcc tttcaagatc accggttctg gtgatggcga gaccccaatt
     1201 gctgaaccgg agaggttatc ggcctcaagt tcatctagaa ggaaatcatt actgggttcc
     1261 ctgataaaac ccttggtcgg aggtggcact ctttcatcca cagtagacaa ccacttgtcg
     1321 acttgatccc gcgccttccc tggctctatt ctcccaggcc gctcagtggg aagcgctggc
     1381 ttctccctct ccttcaggag agtttgagcc aaatttcctg atcgtccttc tatttcatcc
     1441 gtgaagcccc cttgccgaga tcctgctgga acaagaacat ctggggattg tttggtgtac
     1501 agaagtgagg gggcaccgcc gaaaacctcg agagagccgc gagcagttcg agacgcatct
     1561 actttggatg aagtatcagc tggcttacga gcgttgaaag gcttcggagg aacggatggc
     1621 tccgcggacg gcatcatgaa gcctgctttt ttgtacaaac ttgtggagcc tcctgagcct
     1681 ccaagcttgg taccgagctc gaattctgta attgtaaata gtaattgtaa tgttgtttgt
     1741 tgtttgttgt tgttggtaat tgttgaagga tctcaagaag cctgtcctct ccaaatgaaa
     1801 tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg cgtcatccct
     1861 tacgtcagtg gagatatcac atcaatccac ttgctttgaa gacgtggttg gaacgtcttc
     1921 tttttccacg atgttcctcg tgggtggggg tccatctttg ggaccactgt cggtagaggc
     1981 atcttgaacg atagcctttc ctttatcgca atgatggcat ttgtaggagc caccttcctt
     2041 ttctactgtc ctttcgataa gtgacagata gctgggcaat ggaatccgag gaggtttccc
     2101 gatattaccc tttgttgaaa agtctcaata gccctcggtc ttctgagccc gggagatctg
     2161 gcctcgagat cccgggtacc gagctcgaat tctgcggccg cgccctatag tgagtcgtat
     2221 tacactagct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2281 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2341 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgtagtaa
     2401 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     2461 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2521 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2581 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2641 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2701 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2761 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttatgtctgc ttacataaac
     2821 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc aaggccgcga
     2881 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     2941 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     3001 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     3061 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     3121 tggttactca cctctgcgat cccagggaaa acagcattcc aggtattaga agaatatcct
     3181 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     3241 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     3301 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     3361 gttgaacaag tctggaaaga aatgcataag ctattgccat tctcaccgga ttcagtcgtc
     3421 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     3481 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     3541 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     3601 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaattaaaa
     3661 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3721 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3781 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3841 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3901 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3961 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4021 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4081 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4141 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4201 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4261 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4321 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4381 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4441 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4501 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4561 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4621 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     4681 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     4741 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     4801 accatgatta cgccatgcta tttaggtgac actatagaat acgcggccgc aagcttgcat
     4861 gcctgcaggt cgactctaga gcggatccac aattgcggcc gcatctagta acatagatga
     4921 caccgcgcgc gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta
     4981 aatgtataat tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta
     5041 catgttaatt attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag
     5101 accggcaaca ggattcaatc ttaagaaact ttattgccaa atgtttgaac gattctagat
     5161 actagtgcta gcccatgggg agcctcctga gcctccaacc actttgtaca agaaagctgg
     5221 gtctcacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     5281 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact ggtagctcag
     5341 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     5401 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     5461 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     5521 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     5581 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     5641 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     5701 ctgcttcatg tggtcggggt agcgggcgaa gcacatcagg ccgtagccga aggtggtcac
     5761 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     5821 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     5881 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     5941 catggagcct cctgagcctc caaaagcacc atcaaaggag gctgcagttg gatccccatc
     6001 atcccagtcc aaatctttag tctctggttc cttcccaata tggaatactg gagtttctag
     6061 ctttggagga atcatgcaac gaataagagg ccaattgatg tcactgaaaa aggggtgaga
     6121 cttcaaatca ttggctccct tatctgaacc caatcgcttc ttgggatctc tatgcaagag
     6181 accacgtatt acttgctttg caccaatgct cacaggtatt gatgatggaa aaccaagatc
     6241 tttatgcaac acattggcga atgttctttg cctgttacgg cctcggaatg gtgttcgtcc
     6301 aaataacatc tcatacaaga gaatccctag ggcccaccag tccacagagc tgttgtgccc
     6361 gagtccaaga ataatttcag gagcaatgta ttcctcagtc ccgacaaatg aatttgaccg
     6421 tgcagggggt tctgtcagaa ggatcgggcg taatgattcc ttgtttacgc ttctcttacg
     6481 tcgtccagaa ggcacagctg gtttataaag ctggggtcgt gaggcagtca aaaaggatag
     6541 atcaaagtcg gtgagctgta catgtccatc tcgctgtatt aagacgttct caggtttcaa
     6601 gtccctgtac actaccccta cacagtggag atactcaaga gccaatataa tttctgctgc
     6661 ataaaacctt gccgagtcct cttgaaatct cttctgaggt tgtctttcca acaagagaaa
     6721 aagctcacca ccagggcaaa agtctgtgat aagacacaca tgtgttcgtg tctggaaaga
     6781 accataaaga gtgggcaaga aagggtgatc cattttctca agaatctctc tttctgcacg
     6841 agcacgatga accttattac ggttaaccat cacagccttc tccatcgcct tcatggcgta
     6901 aaactcagac gtgtttctta gctcgaccaa atgaacactt cctgtatccc cacaaccaag
     6961 aggtttaatc ggtcgaaagt gattgagtcc cagtttctcg ccagtactca gaaccttgcg
     7021 tacggcatgc catgtagcgc tgttccattt gtgaggcttg ggaagaacaa gacgtgaatg
     7081 gtttgcccac aagtcctcca gactcgtgtt tgcatctggg agctctctca aagctccatc
     7141 aacatttttt gccgtagatt gcacaatttt tttcccttct tcttctgttc tctctgacat
     7201 tcgtcgtctg attggttcaa tgtgctcact gccatcaagt tgaactccga tgaagtactg
     7261 caactctccc ttgttgtcct tcatagcctc aagatgaaaa aggttccaga atggcttccc
     7321 tgttttggtg taatttaaaa gttgcaccgt gatgttttgt tgagatctaa ttgcatcact
     7381 aatcttccga accacaccca tatctgtgtc aggaccttgc agaaacctac aattccgtcc
     7441 aatgatttcc tcacgcgtgt actccgtcaa ctctagaaaa tgatctgatg caaagataat
     7501 gggattgtca ggcagacgtg gatctgtaat aacaaaattc ttctcaattc gctctagagt
     7561 agttgctaag tcc
//
