LOCUS       sequence-367546-        5787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Light-oxygen-voltage-sensing domain 2 linked to RNA binding 
            peptide (on N terminal end) via GS-linker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  AUTHORS   Younas T, Vidallon MLP, Tabor RF, He L
  TITLE     Open-Closed Structure of Light-Responsive Protein LOV2 Regulates Its
            Molecular Interaction with a Binding Partner.
  JOURNAL   J Phys Chem Lett. 2020 Oct 15;11(20):8647-8653. doi: 
            10.1021/acs.jpclett.0c02252. Epub 2020 Sep 29.
  PUBMED    32945680
REFERENCE   2  (bases 1 to 5787)
  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"; doi: 
            "10.1021/acs.jpclett.0c02252"; journalName: "J Phys Chem Lett"; 
            date: "2020-10-15- 15"; volume: "11"; issue: "20"; pages: 
            "8647-8653"
COMMENT     Sequence Label: pET28a-RBP-LOV2
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..48
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     282..299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    314..453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(379..398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     539..561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(555..746)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1555..2637)
                     /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     2599..2618
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2638..2715)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2921..2940
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3024..3043
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3024..3042
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3043..3067
                     /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)."
     RBS             3082..3104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3117..3134
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3147..3167
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3675..3692
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3745..3763)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3759..3806
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3843..4298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3930..3949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4140..4161
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4390..5205)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4390..5205)
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5113..5132
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5197..5216
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(5327..5787,1..128)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
       61 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      121 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      181 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      241 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      301 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
      361 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
      421 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      481 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      541 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
      601 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
      661 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
      721 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
      781 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
      841 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
      901 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
      961 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     1021 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     1081 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     1141 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     1201 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     1261 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     1321 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     1381 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     1441 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     1501 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     1561 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1621 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     1681 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     1741 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     1801 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     1861 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     1921 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     1981 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2041 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2101 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2161 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2221 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2281 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2341 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2401 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2461 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     2521 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     2581 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     2641 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     2701 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     2761 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     2821 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     2881 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     2941 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     3001 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     3061 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atggcacatc
     3121 atcatcatca tcatggcggc ggcagcgaga atctgtattt ccagggcagc tatggtcgta
     3181 aaaagcgtcg tcaacgtcgt cgtccgccgc aaggcggtgg tagcggtggc ggtagcctgg
     3241 cgaccaccct ggagcgtatc gagaagaact tcgtgatcac cgatccgcgt ctgccggata
     3301 acccgatcat tttcgcgagc gatagcttcc tgcaactgac cgaatacagc cgtgaggaaa
     3361 tcctgggtcg taactgccgt tttctgcagg gtccggaaac cgatcgtgcg accgttcgta
     3421 agatccgtga cgcgatcgat aaccagaccg aagtgaccgt tcagctgatt aactacacca
     3481 aaagcggcaa gaaattctgg aacctgttcc acctgcaacc gatgcgtgat cagaagggtg
     3541 atgtgcagta cttcatcggt gtgcaactgg atggcaccga acacgttcgt gatgcggcgg
     3601 aacgtgaggg cgttatgctg attaagaaaa ccgcggaaaa cattgatgaa gcggcgaaag
     3661 aactgtaact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     3721 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3781 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     3841 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3901 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3961 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4021 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4081 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4141 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4201 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4261 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4321 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4381 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     4441 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     4501 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     4561 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     4621 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     4681 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     4741 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     4801 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     4861 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     4921 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     4981 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     5041 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     5101 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     5161 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     5221 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     5281 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5341 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5401 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5461 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5521 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5581 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5641 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5701 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5761 ggtatccggt aagcggcagg gtcggaa
//
