LOCUS       sequence-447072-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The expression of  MBP fused to MpPHOT-LOV1 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  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 6939)
  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-LOV1
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(261..266,267..282,283..289)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    297..313
                     /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     302..324
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             383..1483
                     /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      1340..1349
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1457..1480
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1493..1517
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1851..1875)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1887..1904)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1887..1903)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1896..1918)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2082..2099)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2252..2338
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2430..2457
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2476..2567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2568..2636,2637..3428)
                     /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(2786..2805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3470..3983)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3665..3686)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3877..3896
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4094..4682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4583..4602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4836..4853
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4868..5008
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4934..4953)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5094..5116
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5110..5301)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5797..5874
                     /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             join(5875..6939,1..18)
                     /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(5894..5913)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcaca
       61 attctcatgt ttgacagctt atcatcgact gcacggtgca ccaatgcttc tggcgtcagg
      121 cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg
      181 ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc
      241 aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt
      301 gagcggataa caatttcaca caggaaacag ccagtccgtt taggtgtttt cacgagcact
      361 tcaccaacaa ggaccataga ttatgaaaac tgaagaaggt aaactggtaa tctggattaa
      421 cggcgataaa ggctataacg gtctcgctga agtcggtaag aaattcgaga aagataccgg
      481 aattaaagtc accgttgagc atccggataa actggaagag aaattcccac aggttgcggc
      541 aactggcgat ggccctgaca ttatcttctg ggcacacgac cgctttggtg gctacgctca
      601 atctggcctg ttggctgaaa tcaccccgga caaagcgttc caggacaagc tgtatccgtt
      661 tacctgggat gccgtacgtt acaacggcaa gctgattgct tacccgatcg ctgttgaagc
      721 gttatcgctg atttataaca aagatctgct gccgaacccg ccaaaaacct gggaagagat
      781 cccggcgctg gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca
      841 agaaccgtac ttcacctggc cgctgattgc tgctgacggg ggttatgcgt tcaagtatga
      901 aaacggcaag tacgacatta aagacgtggg cgtggataac gctggcgcga aagcgggtct
      961 gaccttcctg gttgacctga ttaaaaacaa acacatgaat gcagacaccg attactccat
     1021 cgcagaagct gcctttaata aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg
     1081 gtccaacatc gacaccagca aagtgaatta tggtgtaacg gtactgccga ccttcaaggg
     1141 tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa
     1201 caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg actgatgaag gtctggaagc
     1261 ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg aagagttggc
     1321 gaaagatcca cgtattgccg ccaccatgga aaacgcccag aaaggtgaaa tcatgccgaa
     1381 catcccgcag atgtccgctt tctggtatgc cgtgcgtact gcggtgatca acgccgccag
     1441 cggtcgtcag actgtcgatg aagccctgaa agacgcgcag actaattcga tcacaagttt
     1501 gtacaaaaaa gcaggcttcc agcagacctt tgtggtctcg gatgccacac gcacggactt
     1561 ccccattctg tatgccagtg cgggattttt cgccatgaca ggttacactc ccagagaggt
     1621 catcggaaga aactgtcgat ttctccaggg acctgatacc gaccaaagcg acatagagag
     1681 aattcgaact gctctcaagg aggggaagag ttattgtgga agacttctga attacaggaa
     1741 agacggaact cctttctgga accttctgac aattgcacct atcaaggacg acactgggaa
     1801 agtgctcaaa tacattggaa tgcaggtcga ggtcagcaaa tacacatgag acccagcttt
     1861 cttgtacaaa gtggtgatag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     1921 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     1981 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2041 tggcagcttg gctgttttgg cggatgagat aagattttca gcctgataca gattaaatca
     2101 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     2161 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     2221 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2281 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2341 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     2401 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     2461 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2521 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2581 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2641 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2701 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc
     2761 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     2821 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2881 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2941 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3001 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3061 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3121 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3181 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3241 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3301 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3361 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3421 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta ccccggttga
     3481 taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaaacgt
     3541 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3601 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag ggttgagtgt
     3661 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3721 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     3781 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3841 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3901 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3961 taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt gataatctca
     4021 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4081 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4141 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4201 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4261 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4321 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4381 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4441 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4501 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4561 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4621 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4681 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4741 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4801 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4861 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4921 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     4981 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     5041 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5101 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     5161 ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     5221 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5281 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     5341 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     5401 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     5461 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     5521 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     5581 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     5641 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     5701 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     5761 cacccgtggc caggacccaa cgctgcccga aattccgaca ccatcgaatg gtgcaaaacc
     5821 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     5881 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     5941 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     6001 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     6061 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     6121 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     6181 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     6241 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     6301 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     6361 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     6421 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     6481 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     6541 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     6601 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     6661 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     6721 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     6781 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     6841 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6901 gccgattcat taatgcagct ggcacgacag gtttcccga
//
