LOCUS       sequence-448719-        7011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The expression of MBP fused to MpPHOT-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-MpPHOT-LOV2
FEATURES             Location/Qualifiers
     source          1..7011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      21..48
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        67..158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(159..227,228..1019)
                     /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     complement(377..396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1061..1574)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1256..1277)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1468..1487
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1685..2273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2174..2193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2459..2599
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2525..2544)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2685..2707
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2701..2892)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3388..3465
                     /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             3466..4548
                     /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     complement(3485..3504)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4791..4796,4797..4812,4813..4819)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4827..4843
                     /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     4832..4854
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4913..6013
                     /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"
     regulatory      5870..5879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     5987..6010
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    6023..6047
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6453..6477)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6489..6506)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6489..6505)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6498..6520)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6684..6701)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6854..6940
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
       61 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      121 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      181 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      241 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      301 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttc ccaatgatga
      361 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
      421 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
      481 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
      541 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
      601 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
      661 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
      721 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
      781 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
      841 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
      901 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
      961 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1021 tgtcagacca agtttactca tatatacttt agattgattt accccggttg ataatcagaa
     1081 aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaaacg ttaatatttt
     1141 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1201 cggcaaaatc ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt
     1261 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1321 ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag
     1381 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1441 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1501 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1561 gctacagggc gcgtaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1621 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1681 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1741 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1801 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     1861 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1921 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1981 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2041 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2101 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2161 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2221 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2281 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2341 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2401 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2461 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     2521 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     2581 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2641 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2701 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     2761 gtggtcgtgc agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     2821 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     2881 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     2941 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     3001 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     3061 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     3121 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     3181 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     3241 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     3301 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     3361 ccaggaccca acgctgcccg aaattccgac accatcgaat ggtgcaaaac ctttcgcggt
     3421 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     3481 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     3541 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     3601 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     3661 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     3721 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     3781 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     3841 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     3901 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     3961 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     4021 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     4081 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     4141 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     4201 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     4261 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     4321 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     4381 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     4441 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4501 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     4561 taatgtaagt tagctcactc attaggcaca attctcatgt ttgacagctt atcatcgact
     4621 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
     4681 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     4741 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     4801 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
     4861 ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga ttatgaaaac
     4921 tgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga
     4981 agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa
     5041 actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg
     5101 ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga
     5161 caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa
     5221 gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct
     5281 gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa
     5341 aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc
     5401 tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg
     5461 cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa
     5521 acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac
     5581 agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta
     5641 tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct
     5701 gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa
     5761 ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt
     5821 agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga
     5881 aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc
     5941 cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa
     6001 agacgcgcag actaattcga tcacaagttt gtacaaaaaa gcaggcttcg agagtattga
     6061 tgacagtgag cgagccaagg aaattcgtag gggaatggat ttggccacaa cgttagagcg
     6121 aattgagaag aatttcgtga ttacagatcc tcgattgccg gataacccca ttattttcgc
     6181 atccgaccac ttcttagagt taacggagta tactcgagaa gaaattatcg gaagaaattg
     6241 tcggtttctg caagggcccg atacggacat ggatgttgtt cggaaaatta gtgatgctat
     6301 caaacaacag caaaacatca ctgttcaact tctgaattac accaaaggcg gaaaaccgtt
     6361 ttggaatttg tttcacttgg aagcaatgaa ggacaacaag ggtgagttgc aatattttat
     6421 cggagtccag ctcgatggaa gtgagcactg agacccagct ttcttgtaca aagtggtgat
     6481 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     6541 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     6601 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcagct tggctgtttt
     6661 ggcggatgag ataagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     6721 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     6781 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     6841 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     6901 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     6961 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa c
//
