LOCUS       sequence-326604-        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 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: pQE-BR1
FEATURES             Location/Qualifiers
     source          1..5054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..33
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(57..71,72..77,78..94,95..101)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    77..93
                     /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    109..125
                     /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             144..155
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             174..191
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      558..652
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             696..1355
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(762..781)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1420..1506
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1599..2681)
                     /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     2643..2662
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2682..2759)
                     /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(2843..2865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2951..3091
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3006..3025
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3106..3123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3277..3865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3357..3376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4036..4827,4828..4896))
                     /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     4659..4678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4897..5001)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagaaatcat
       61 aaaaaattta tttgctttgt gagcggataa caattataat agattcaatt gtgagcggat
      121 aacaatttca cacagaattc attaaagagg agaaattaac tatgagagga tcgcatcacc
      181 atcaccatca cggatccatg attgagaaaa attatgtcat caccgatccc cggttgcctg
      241 ataacccaat aattttcgcc tctgatggtt ttctggaact gacaggatat agtcgtgaag
      301 aaatacttgg ccgtaatgcg cgctttctcc aaggtccgga gaccgatcag gcaaccgtac
      361 agaagatccg tgacgctatc cgcgaccaaa gagaaacgac tgtgcaactg attaactaca
      421 cgaaatctgg cagaaagttt tggaatctgc tccatctgca gccggttcgc gatcagaagg
      481 gcgagttgca gtatttcatc ggagttcagc tggacggaag tgatcgcgtt tgataaaagc
      541 ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag aactccatct
      601 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta
      661 gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
      721 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
      781 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
      841 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
      901 ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
      961 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     1021 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     1081 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     1141 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     1201 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     1261 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     1321 aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     1381 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     1441 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     1501 acaaatccgc cctctagatt acgtgcagtc gatgataagc tgtcaaacat gagaattgtg
     1561 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1621 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1681 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     1741 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     1801 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     1861 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     1921 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     1981 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     2041 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     2101 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     2161 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     2221 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     2281 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     2341 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     2401 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     2461 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     2521 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     2581 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     2641 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     2701 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg
     2761 gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc tgcctcgcgc
     2821 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2881 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2941 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     3001 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     3061 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     3121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5041 cattattatc atga
//
