LOCUS       sequence-326603-        5054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes an engineered oxygen-independent flavin binding
            fluorescent protein with N-terminal 6xHis tag. Its thermostability 
            and quantum yield is improved compared to wild type..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5054)
  AUTHORS   Ko S, Hwang B, Na JH, Lee J, Jung ST
  TITLE     Engineered Arabidopsis Blue Light Receptor LOV Domain Variants with 
            Improved Quantum Yield, Brightness, and Thermostability.
  JOURNAL   J Agric Food Chem. 2019 Oct 30;67(43):12037-12043. doi: 
            10.1021/acs.jafc.9b05473. Epub 2019 Oct 17.
  PUBMED    31581772
REFERENCE   2  (bases 1 to 5054)
  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.jafc.9b05473"; journalName: "J Agric Food Chem"; date: 
            "2019-10-30- 30"; volume: "67"; issue: "43"; pages: "12037-12043"
COMMENT     Sequence Label: pQE80L-SH3
FEATURES             Location/Qualifiers
     source          1..5054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..34
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(58..72,73..78,79..95,96..102)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    78..94
                     /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)."
     protein_bind    110..126
                     /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             145..156
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             175..192
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      559..653
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             697..1356
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(763..782)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1421..1507
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1600..2682)
                     /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     2644..2663
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2683..2760)
                     /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."
     primer_bind     complement(2844..2866)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2952..3092
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3007..3026
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3107..3124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3278..3866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3358..3377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4037..4828,4829..4897))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4660..4679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4898..5002)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacct cgagaaatca
       61 taaaaaattt atttgctttg tgagcggata acaattataa tagattcaat tgtgagcgga
      121 taacaatttc acacagaatt cattaaagag gagaaattaa ctatgagagg atcgcatcac
      181 catcaccatc acggatccat gattgagaaa aattatgtca tcaccgatcc ccggttgcct
      241 gataacccaa taattttcgc ctcagatggt tttctggaac tgacagaata tagtcgtgaa
      301 gaaatacttg gccgtaatgc gcgctttctc caaggtccgg agaccgatca ggcaaccgta
      361 cagaagatcc gtgacgctat ccgcgaccaa agagaaacga ctgtgcaact gattaactac
      421 acgaaatctg gcaaagaatt ttggaatctg ttccatctgc agccggttcg caatcagaag
      481 ggcgagttgc agtatttcat cggggttcag caggacggaa gcgatcacgt gtgataaaag
      541 cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
      601 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct
      661 agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
      721 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
      781 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
      841 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
      901 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
      961 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     1021 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     1081 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     1141 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     1201 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     1261 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     1321 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     1381 ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac gaaaggctca
     1441 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1501 gacaaatccg ccctctagat tacgtgcagt cgatgataag ctgtcaaaca tgagaattgt
     1561 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1621 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1681 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     1741 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     1801 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     1861 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     1921 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     1981 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     2041 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     2101 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     2161 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     2221 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     2281 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     2341 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     2401 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     2461 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     2521 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     2581 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     2641 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     2701 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc
     2761 ggaatttcgg gcagcgttgg gtcctggcca cgggtgcgca tgatctagag ctgcctcgcg
     2821 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     2881 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     2941 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     3001 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     3061 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     3121 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3181 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3241 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3301 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3361 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3421 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3481 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3541 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3601 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3661 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3721 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3781 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3841 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3901 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3961 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4021 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4081 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4141 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4201 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4261 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4321 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4381 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4441 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4501 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4561 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4621 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4681 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4741 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4801 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4861 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4921 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4981 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5041 ccattattat catg
//
