LOCUS       sequence-447064-        6931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of MpPHOT in plant cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6931)
  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 6931)
  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
FEATURES             Location/Qualifiers
     source          1..6931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     protein_bind    complement(6310..6334)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6379..6629
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(6694..6712)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6695..6712)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6730..6746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6730..6746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6743..6765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6754..6770
                     /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(6778..6784,6785..6802,6803..6808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6823..6844
                     /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 cctccacgga ttcatcgtct gcgaagcctt
     3001 acaatctgcg caagcaaggg gacacgccca caaaacccga cgtgggtcga gacgctcgag
     3061 gctctctcga agtattcggc ggggcgccac aaacctcagc attgctgttc tcgaggtcag
     3121 ctgcggaaga tgccttggca gcggccggaa ttcgaccctc gcaaattctg agcccctcgg
     3181 gctccggcag gcttctgccc gcgctcggcc cgttagggtc ggacggacag gcgctgccgc
     3241 tgtcgcagtc catgctcaag gagcgcgaga agcaagttct gggcacagag cgcggcgtga
     3301 gaacggactc gggacgcgcc agggatcaag tggacaggtg gctggctact gtcgacgagc
     3361 gcgctccgcc ttccaaaggc tcggccaagg agccgagcac ggatttctcg ttggacgacc
     3421 tggagctgcc atcttccggc gctgctgctg gcccaggagc cagaggcgca gcagccttgg
     3481 cctcgcccga atgggctggg gccaaaggca aagctgtggt cgagaaggag ccgagcgaag
     3541 tggccggagg aggcttccag gcgcccgaag gtgtgtttcc cctggggggc gacaaggtgg
     3601 tgcccatggt gtcgggcccg tcggacttgt cagaggaggt gcaggccgag cgagccgcgc
     3661 aatggggagt gctgatgaag agcacggcag atctggcctc gaccggccgc gcgtccgggt
     3721 ccaaggacga gcggatttcg gcctccggca tgtcccgcag aacttcggaa ggatccgaag
     3781 gcgaggttcc tcgtgtctct agagatttga aagatgctct cgcttctttc cagcagacct
     3841 ttgtggtctc ggatgccaca cgcacggact tccccattct gtatgccagt gcgggatttt
     3901 tcgccatgac aggttacact cccagagagg tcatcggaag aaactgtcga tttctccagg
     3961 gacctgatac cgaccaaagc gacatagaga gaattcgaac tgctctcaag gaggggaaga
     4021 gttattgtgg aagacttctg aattacagga aagacggaac tcctttctgg aaccttctga
     4081 caattgcacc tatcaaggac gacactggga aagtgctcaa atacattgga atgcaggtcg
     4141 aggtcagcaa atacacagaa ggaaacaagt ccaatgaagt tcggccgaac ggaatgccgg
     4201 cttcacttat caaatacgac gctcggcagc aggacagagc cacgagttct gttcaggaac
     4261 tggtgggagc tttgaaacat cctcaccaat caccgcaaga taagccctcg gaaggaggca
     4321 tgcaatcgat gttttcagtt cctccattga aagacgaact gcagttgaag gtaccctcac
     4381 gcttttcaac gcagtcggtt gtcgcactgc cgctcgaggg tagccaatcg gcgaaagcaa
     4441 gggcaaaaag tgagtcagtg agtcgactga ccgatgtccc agaacaacct ggaatggctg
     4501 atggcggcct tcgagggggg cagagccacc gcagatcgtc ggggttgttg caaatgctca
     4561 ggaaaccgaa atcacaacag ccagaagctg atgtcgtcga catcgatgac ttggaccttg
     4621 acgatgatgc cccacagacc atggatgaga ggccagagag tattgatgac agtgagcgag
     4681 ccaaggaaat tcgtagggga atggatttgg ccacaacgtt agagcgaatt gagaagaatt
     4741 tcgtgattac agatcctcga ttgccggata accccattat tttcgcatcc gaccacttct
     4801 tagagttaac ggagtatact cgagaagaaa ttatcggaag aaattgtcgg tttctgcaag
     4861 ggcccgatac ggacatggat gttgttcgga aaattagtga tgctatcaaa caacagcaaa
     4921 acatcactgt tcaacttctg aattacacca aaggcggaaa accgttttgg aatttgtttc
     4981 acttggaagc aatgaaggac aacaagggtg agttgcaata ttttatcgga gtccagctcg
     5041 atggaagtga gcacatcgaa cccatcaggc gacgactctc cgaaaagact gaagaggaag
     5101 gcaaaaggat tgtgcaagct acagcgaaga atgttgacgg agcagtgagg gagcttccag
     5161 atgcgaatat gagcatcgag gatttgtggg cgaatcactc tcgtgttgtc tttcctaggc
     5221 ctcacaaacg tcagagttcc acatggcatg ccatgcgaaa gatactcagt tctggagaca
     5281 aattgggtct caaccacttc aggccaatca aacctcttgg ttgtggagat accggcagtg
     5341 tacatctggt ggagctgaga gggacgagtg agttctatgc catgaaagcc atggataaga
     5401 atgttatggt gaatcggaat aaggttcacc gtgctcgggc agagagagaa attctggaga
     5461 aaatggatca tccgtttttg cccacgttat atggctcctt ccagactagg acacatgtgt
     5521 gtctcatcac cgatttttgt cctggtggtg agctgttctt gcttttggaa agacaacccc
     5581 agaagaagtt ccgtgaggag tctgctaggt tttatgcagc cgagattgtt cttgccctcg
     5641 aatatcttca ctgcatgggt gttgtataca gggacctgaa acctgaaaac gttctggtcc
     5701 aaaaagatgg gcatgtgcag ctcactgatt tcgacctttc cttcttgact gcctctcgtc
     5761 ctcagctgct gaaaccagtc gtaccatctg gccgaaagaa tagacgagca agggagaacc
     5821 tacgaccaat cctccttaca gagccaatag caaagtctaa ctccttcgtt ggaactgagg
     5881 agtatattgc gccggaaatt atccttggtt tgggacacaa cagctctgta gattggtggg
     5941 ctctaggtat tcttctgtac gagatgttat ttggacgaac accctttaga ggacgcaacc
     6001 ggcaaaggac tttcgcaaat gttctgcata aagatcttgc cttcccaagc tccatacctg
     6061 taagcttggc tgccaaacaa gttattcgtg ggctgctaca gagagatccc aagaagaggt
     6121 tggggtcaga caagggagcc cacgacttaa aacaacatca attcttccgg ggcatcaatt
     6181 ggcctctcat ccgctgcatg actcctccac cgctagaaac accaatctac caaatcggca
     6241 aagaagcgga ctccaaagac ctggactggg aggagttgga tcccagtgcc gcctcatttg
     6301 atgaatatga cccagctttc ttgtacaaag tggttggagg ctcaggaggc tccccatggg
     6361 ctagcactag tatctagaat cgttcaaaca tttggcaata aagtttctta agattgaatc
     6421 ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa
     6481 taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc
     6541 aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat
     6601 cgcgcgcggt gtcatctatg ttactagatg cggccgcaat tgtggatccg ctctagagtc
     6661 gacctgcagg catgcaagct tgcggccgcg tattctatag tgtcacctaa atagcatggc
     6721 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6781 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6901 ttaatgaatc ggccaacgcg cggggagagg c
//
