LOCUS       sequence-447071-        6930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The expression of  MBP fused to AePHOT-LOV1 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6930)
  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 6930)
  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-LOV1
FEATURES             Location/Qualifiers
     source          1..6930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      48..75
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        94..185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(186..254,255..1046)
                     /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(404..423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1088..1601)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1283..1304)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1495..1514
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1712..2300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2201..2220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2454..2471
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2486..2626
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2552..2571)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2712..2734
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2728..2919)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3415..3492
                     /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             3493..4575
                     /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(3512..3531)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4818..4823,4824..4839,4840..4846)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4854..4870
                     /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     4859..4881
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             4940..6040
                     /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      5897..5906
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     6014..6037
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    6050..6074
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6399..6423)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6435..6452)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6435..6451)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6444..6466)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6630..6647)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6800..6886
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
       61 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
      121 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      181 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      241 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      301 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      361 gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      421 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      481 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      541 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      601 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      661 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      721 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      781 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      841 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      901 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      961 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1021 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1081 ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     1141 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     1201 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc
     1261 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     1321 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     1381 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     1441 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     1501 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1561 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga
     1621 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1681 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1741 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1801 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1861 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1921 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1981 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2041 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2101 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2161 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2221 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2281 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2341 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2401 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2461 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     2521 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2581 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     2641 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2701 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2761 gaggcagctg cggtaaagct catcagcgtg gtcgtgcagc gattcacaga tgtctgcctg
     2821 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     2881 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     2941 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     3001 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     3061 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     3121 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     3181 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     3241 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     3301 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     3361 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaaa ttccgacacc
     3421 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     3481 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     3541 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     3601 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     3661 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     3721 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     3781 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     3841 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     3901 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     3961 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     4021 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     4081 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     4141 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     4201 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     4261 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     4321 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     4381 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     4441 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     4501 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     4561 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacaatt
     4621 ctcatgtttg acagcttatc atcgactgca cggtgcacca atgcttctgg cgtcaggcag
     4681 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     4741 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     4801 tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     4861 cggataacaa tttcacacag gaaacagcca gtccgtttag gtgttttcac gagcacttca
     4921 ccaacaagga ccatagatta tgaaaactga agaaggtaaa ctggtaatct ggattaacgg
     4981 cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat
     5041 taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac
     5101 tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc
     5161 tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac
     5221 ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt
     5281 atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc
     5341 ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga
     5401 accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa
     5461 cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac
     5521 cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc
     5581 agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc
     5641 caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca
     5701 accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa
     5761 agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt
     5821 taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggcgaa
     5881 agatccacgt attgccgcca ccatggaaaa cgcccagaaa ggtgaaatca tgccgaacat
     5941 cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg
     6001 tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgatca caagtttgta
     6061 caaaaaagca ggcttcacct ttgtagtgtc tgatgctacc cgtcctgact acccaatcct
     6121 ctatgccagt gcggggttct tcgacatgac aggatatgca ccgaaagagg tcatcgggag
     6181 gaactgtcgg tttctccagg gaccggaaac tgatcaagaa gatattgaga ggattcgatc
     6241 tgcactgaag gatgggaaga ctttttgtgg gagacttcta aattacagaa agaatggaac
     6301 ccctttctgg aatctcctta caatagcacc cattaaagac gacaatggga agattctgaa
     6361 atacattgga atgcaggtgg aagttagcaa gtatactgac ccagctttct tgtacaaagt
     6421 ggtgatagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     6481 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     6541 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcagcttggc
     6601 tgttttggcg gatgagataa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     6661 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     6721 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     6781 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     6841 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     6901 tttgaacgtt gcgaagcaac ggcccggagg
//
