LOCUS       sequence-448720-        7011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The expression of  MBP fused to AePHOT-LOV2 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7011)
  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 7011)
  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: pKM596-AePHOT-LOV2
FEATURES             Location/Qualifiers
     source          1..7011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..127
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     primer_bind     291..313
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     305..322
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     306..322
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    334..358
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(764..788)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(798..1898)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(801..824)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     regulatory      932..941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1957..1979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1968..1984
                     /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(1992..1998,1999..2014,2015..2020))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(2263..3345)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3307..3326
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3346..3423)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3919..4110
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4104..4126)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4212..4352
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4267..4286
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4367..4384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4538..5126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4618..4637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      5237..5750
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5324..5343)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5534..5555
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5792..6583,6584..6652))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6415..6434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6653..6744)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(6763..6790)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6882..6968)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 accatcggcg ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc
       61 gctactgccg ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg
      121 tatcaggctg aaaatcttat ctcatccgcc aaaacagcca agctgccatt cgccattcag
      181 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      241 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      301 acgttgtaaa acgacggcca gtgccaagct atcaccactt tgtacaagaa agctgggtca
      361 atgtgctcac tgccatcaag ttgaactccg atgaagtact gcaactctcc cttgttgtcc
      421 ttcatagcct caagatgaaa aaggttccag aatggcttcc ctgttttggt gtaatttaaa
      481 agttgcaccg tgatgttttg ttgagatcta attgcatcac taatcttccg aaccacaccc
      541 atatctgtgt caggaccttg cagaaaccta caattccgtc caatgatttc ctcacgcgtg
      601 tactccgtca actctagaaa atgatctgat gcaaagataa tgggattgtc aggcagacgt
      661 ggatctgtaa taacaaaatt cttctcaatt cgctctagag tagttgctaa gtccatgcct
      721 cgacgaatct ccgtagcccg ctctgaatta tccaaacttt cgaagcctgc ttttttgtac
      781 aaacttgtga tcgaattagt ctgcgcgtct ttcagggctt catcgacagt ctgacgaccg
      841 ctggcggcgt tgatcaccgc agtacgcacg gcataccaga aagcggacat ctgcgggatg
      901 ttcggcatga tttcaccttt ctgggcgttt tccatggtgg cggcaatacg tggatctttc
      961 gccaactctt cctcgtaaga cttcagcgct acggcaccca gcggtttgtc tttattaacc
     1021 gcttccagac cttcatcagt cagcagatag ttttcgagga actcttttgc cagctctttg
     1081 ttcggactgg cggcgttaat acctgcgctc agcacgccaa cgaacggttt ggatggttga
     1141 cccttgaagg tcggcagtac cgttacacca taattcactt tgctggtgtc gatgttggac
     1201 catgcccacg ggccgttgat ggtcatcgct gtttcgcctt tattaaaggc agcttctgcg
     1261 atggagtaat cggtgtctgc attcatgtgt ttgtttttaa tcaggtcaac caggaaggtc
     1321 agacccgctt tcgcgccagc gttatccacg cccacgtctt taatgtcgta cttgccgttt
     1381 tcatacttga acgcataacc cccgtcagca gcaatcagcg gccaggtgaa gtacggttct
     1441 tgcaggttga acatcagcgc gctcttacct ttcgctttca gttctttatc cagcgccggg
     1501 atctcttccc aggtttttgg cgggttcggc agcagatctt tgttataaat cagcgataac
     1561 gcttcaacag cgatcgggta agcaatcagc ttgccgttgt aacgtacggc atcccaggta
     1621 aacggataca gcttgtcctg gaacgctttg tccggggtga tttcagccaa caggccagat
     1681 tgagcgtagc caccaaagcg gtcgtgtgcc cagaagataa tgtcagggcc atcgccagtt
     1741 gccgcaacct gtgggaattt ctcttccagt ttatccggat gctcaacggt gactttaatt
     1801 ccggtatctt tctcgaattt cttaccgact tcagcgagac cgttatagcc tttatcgccg
     1861 ttaatccaga ttaccagttt accttcttca gttttcataa tctatggtcc ttgttggtga
     1921 agtgctcgtg aaaacaccta aacggactgg ctgtttcctg tgtgaaattg ttatccgctc
     1981 acaattccac acattatacg agccgatgat taattgtcaa cagctcattt cagaatattt
     2041 gccagaaccg ttatgatgtc ggcgcaaaaa acattatcca gaacgggagt gcgccttgag
     2101 cgacacgaat tatgcagtga tttacgacct gcacagccat accacagctt ccgatggctg
     2161 cctgacgcca gaagcattgg tgcaccgtgc agtcgatgat aagctgtcaa acatgagaat
     2221 tgtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     2281 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2341 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     2401 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     2461 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     2521 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     2581 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     2641 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     2701 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     2761 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     2821 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     2881 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     2941 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     3001 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     3061 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     3121 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     3181 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     3241 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     3301 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     3361 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     3421 gtcggaattt cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     3481 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     3541 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     3601 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     3661 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     3721 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     3781 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     3841 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     3901 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     3961 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     4021 gcagacatct gtgaatcgct gcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     4081 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4141 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4201 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     4261 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     4321 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     4381 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4441 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4501 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4561 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4621 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4681 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4741 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4801 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4861 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4921 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4981 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5041 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5101 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5161 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5221 tcacctagat ccttttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5281 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5341 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5401 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     5461 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5521 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     5581 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5641 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttaaatattt
     5701 gcttatacaa tcttcctgtt tttggggctt ttctgattat caaccggggt aaatcaatct
     5761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     6421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg gaaacgttct tcggggcgaa
     6481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6721 aatgtattta gaaaaataaa caaaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     6781 atggccttct gcttaatttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     6841 ccgggccgtt gcttcgcaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     6901 gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     6961 tttatttgat gcctggcagt tccctactct cgcatgggga gaccccacac t
//
