LOCUS       sequence-447059-        6838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of AePHOT in plant cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  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 6838)
  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
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..47)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(201..789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(281..300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(947..1762)
                     /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     1670..1689
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1754..1773
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1947..1965
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2003..2025)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2125..2144
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2338..2360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2352..2369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2353..2369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2379..2398
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2379..2397
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2474..2818
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2761..2781
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    2838..2893
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    2937..2961
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3168..3179
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(6217..6241)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6286..6536
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(6601..6619)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6602..6619)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6637..6653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6637..6653)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6650..6672)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6661..6677
                     /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(6685..6691,6692..6709,6710..6715))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6730..6751
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
       61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      901 aattaaaaat gaagttttaa atcaatctaa agtatatatg ttttaattag aaaaactcat
      961 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     1021 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     1081 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     1141 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     1201 atggcaatag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     1261 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     1321 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     1381 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     1441 ggaatgctgt tttccctggg atcgcagagg tgagtaacca tgcatcatca ggagtacgga
     1501 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     1561 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     1621 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     1681 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctt gagcaagacg
     1741 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacataa
     1801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1921 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1981 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2041 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2101 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2161 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ttactacagg cgccattcgc
     2221 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2281 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2341 agtcacgacg ttgtaaaacg acggccagtg agctagtgta atacgactca ctatagggcg
     2401 cggccgcaga attcgagctc ggtacccggg atctcgaggc cagatctccc gggctcagaa
     2461 gaccgagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc ctcggattcc
     2521 attgcccagc tatctgtcac ttatcgaaag gacagtagaa aaggaaggtg gctcctacaa
     2581 atgccatcat tgcgataaag gaaaggctat cgttcaagat gcctctaccg acagtggtcc
     2641 caaagatgga cccccaccca cgaggaacat cgtggaaaaa gaagacgttc caaccacgtc
     2701 ttcaaagcaa gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca
     2761 ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg agaggacagg
     2821 cttcttgaga tccttcaaca attaccaaca acaacaaaca acaaacaaca ttacaattac
     2881 tatttacaat tacagaattc gagctcggta ccaagcttgg aggctcagga ggctccacaa
     2941 gtttgtacaa aaaagcaggc ttcatgatgc cgtccgcgga gccatccgtt cctccgaagc
     3001 ctttcaacgc tcgtaagcca gctgatactt catccaaagt agatgcgtct cgaactgctc
     3061 gcggctctct cgaggttttc ggcggtgccc cctcacttct gtacaccaaa caatccccag
     3121 atgttcttgt tccagcagga tctcggcaag ggggcttcac ggatgaaata gaaggacgat
     3181 caggaaattt ggctcaaact ctcctgaagg agagggagaa gccagcgctt cccactgagc
     3241 ggcctgggag aatagagcca gggaaggcgc gggatcaagt cgacaagtgg ttgtctactg
     3301 tggatgaaag agtgccacct ccgaccaagg gttttatcag ggaacccagt aatgatttcc
     3361 ttctagatga acttgaggcc gataacctct ccggttcagc aattggggtc tcgccatcac
     3421 cagaaccggt gatcttgaaa ggcaaacgtg tcgggaagga ttctgaattg gggaacggag
     3481 caggagcact cgtgcaatca gtaatacccg atccttccga tgaggcaagt tcagagagag
     3541 gattgcaatg gccagcagca ttgaaagcag tagtagcagc agatttgact gggcgtgcct
     3601 caggctcccg agacgatcgg atatcgggat tgtcagggag gacttcggaa gcatctgatg
     3661 gagggagtga ttcgcaaact gtccctcgca tttctcggga tctcaaggat gcccttgctt
     3721 cgttccagca aacctttgta gtgtctgatg ctacccgtcc tgactaccca atcctctatg
     3781 ccagtgcggg gttcttcgac atgacaggat atgcaccgaa agaggtcatc gggaggaact
     3841 gtcggtttct ccagggaccg gaaactgatc aagaagatat tgagaggatt cgatctgcac
     3901 tgaaggatgg gaagactttt tgtgggagac ttctaaatta cagaaagaat ggaacccctt
     3961 tctggaatct ccttacaata gcacccatta aagacgacaa tgggaagatt ctgaaataca
     4021 ttggaatgca ggtggaagtt agcaagtata ctgaaggaaa tagggcaaac gatgtccgac
     4081 ccaatggaat gccagcatcc ctgataaaat atgatgctcg gcagcaagac agggcggcaa
     4141 gttctgttca agaactggtc ggtgctctca aacatccaaa tggtaatgga caccaagcat
     4201 cacctccacg aaaaagcact ggtggcggtg ctccaccact gttttcattt ccacccttga
     4261 aagatgagct acagctgaag ttaccttccc gctattcaac tcaatcttct gcaatgggga
     4321 agattcctga ggactataat cagccgaaat ttagatcaaa aagtgaatca gtgaacagaa
     4381 ctccagatga agcagaccca cgttcagtgg ggggaaatgc tgggaagagc caaagcactc
     4441 gtcggccttc cggactgatg cagttgatta agaaaccagc aaagcctgca ccacctccac
     4501 cagaagtaga tgatcttgat ctagatgatt ctgtaccgaa tctggaggac agagaaagtt
     4561 tggataattc agagcgggct acggagattc gtcgaggcat ggacttagca actactctag
     4621 agcgaattga gaagaatttt gttattacag atccacgtct gcctgacaat cccattatct
     4681 ttgcatcaga tcattttcta gagttgacgg agtacacgcg tgaggaaatc attggacgga
     4741 attgtaggtt tctgcaaggt cctgacacag atatgggtgt ggttcggaag attagtgatg
     4801 caattagatc tcaacaaaac atcacggtgc aacttttaaa ttacaccaaa acagggaagc
     4861 cattctggaa cctttttcat cttgaggcta tgaaggacaa caagggagag ttgcagtact
     4921 tcatcggagt tcaacttgat ggcagtgagc acattgaacc aatcagacga cgaatgtcag
     4981 agagaacaga agaagaaggg aaaaaaattg tgcaatctac ggcaaaaaat gttgatggag
     5041 ctttgagaga gctcccagat gcaaacacga gtctggagga cttgtgggca aaccattcac
     5101 gtcttgttct tcccaagcct cacaaatgga acagcgctac atggcatgcc gtacgcaagg
     5161 ttctgagtac tggcgagaaa ctgggactca atcactttcg accgattaaa cctcttggtt
     5221 gtggggatac aggaagtgtt catttggtcg agctaagaaa cacgtctgag ttttacgcca
     5281 tgaaggcgat ggagaaggct gtgatggtta accgtaataa ggttcatcgt gctcgtgcag
     5341 aaagagagat tcttgagaaa atggatcacc ctttcttgcc cactctttat ggttctttcc
     5401 agacacgaac acatgtgtgt cttatcacag acttttgccc tggtggtgag ctttttctct
     5461 tgttggaaag acaacctcag aagagatttc aagaggactc ggcaaggttt tatgcagcag
     5521 aaattatatt ggctcttgag tatctccact gtgtaggggt agtgtacagg gacttgaaac
     5581 ctgagaacgt cttaatacag cgagatggac atgtacagct caccgacttt gatctatcct
     5641 ttttgactgc ctcacgaccc cagctttata aaccagctgt gccttctgga cgacgtaaga
     5701 gaagcgtaaa caaggaatca ttacgcccga tccttctgac agaaccccct gcacggtcaa
     5761 attcatttgt cgggactgag gaatacattg ctcctgaaat tattcttgga ctcgggcaca
     5821 acagctctgt ggactggtgg gccctaggga ttctcttgta tgagatgtta tttggacgaa
     5881 caccattccg aggccgtaac aggcaaagaa cattcgccaa tgtgttgcat aaagatcttg
     5941 gttttccatc atcaatacct gtgagcattg gtgcaaagca agtaatacgt ggtctcttgc
     6001 atagagatcc caagaagcga ttgggttcag ataagggagc caatgatttg aagtctcacc
     6061 cctttttcag tgacatcaat tggcctctta ttcgttgcat gattcctcca aagctagaaa
     6121 ctccagtatt ccatattggg aaggaaccag agactaaaga tttggactgg gatgatgggg
     6181 atccaactgc agcctccttt gatggtgctt tttaagaccc agctttcttg tacaaagtgg
     6241 ttggaggctc aggaggctcc ccatgggcta gcactagtat ctagaatcgt tcaaacattt
     6301 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     6361 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     6421 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     6481 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatgcgg
     6541 ccgcaattgt ggatccgctc tagagtcgac ctgcaggcat gcaagcttgc ggccgcgtat
     6601 tctatagtgt cacctaaata gcatggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6661 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6721 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6781 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggc
//
