LOCUS       sequence-255641-        6158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reporter protein which emits bright blue fluorescence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6158)
  AUTHORS   Wu N, Kobayashi N, Tsuda K, Unzai S, Saotome T, Kuroda Y, Yamazaki T
  TITLE     Solution structure of Gaussia Luciferase with five disulfide bonds 
            and identification of a putative coelenterazine binding cavity by 
            heteronuclear NMR.
  JOURNAL   Sci Rep. 2020 Nov 18;10(1):20069. doi: 10.1038/s41598-020-76486-4.
  PUBMED    33208800
REFERENCE   2  (bases 1 to 6158)
  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: "Sci Rep."; 
            date: "2020-11-18"; volume: "10(1):20069. doi"; pages: " 
            10.1038/s41598-020-76486-4"
COMMENT     Sequence Label: pET21c-His-Xa-Gluc-Xa-C9D-TGinsert
FEATURES             Location/Qualifiers
     source          1..6158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             73..90
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             91..102
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             613..624
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(632..661)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(635..664)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     804..824
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             871..888
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(941..959)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      955..1002
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1039..1494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1126..1145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1336..1357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1520..1624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1625..1693,1694..2485)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1843..1862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2656..3244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3145..3164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3398..3415
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3430..3569
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3495..3514)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3655..3677
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3671..3862)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4671..5753)
                     /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     5715..5734
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5754..5831)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6037..6056
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6140..6158,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6140..6158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tgcatcacca tcaccatcac atcgaaggta ggactggcaa accaactgaa
      121 aacaatgaag atttcaacat tgtagctgta gctagcaact ttgctacaac ggatctcgat
      181 gctgaccgtg gtaaattgcc cggaaaaaaa ttaccacttg aggtactcaa agaaatggaa
      241 gccaatgcta ggaaagctgg ctgcactagg ggatgtctga tatgcctgtc acacatcaag
      301 tgtacaccca aaatgaagaa gtttatccca ggaagatgcc acacctatga aggagacaaa
      361 gaaagtgcac agggaggaat aggagaggct attgttgaca ttcctgcaat tcctcggttt
      421 aaggatttgg aaccgatgga acaattcatt gcacaagttg acctatgtgt agactgcaca
      481 actggatgcc tcaaaggtct tgccaatgtg caatgttctg atttactcaa gaaatggctg
      541 ccacaaagat gtgcaacttt tgctagcaaa attcaaggcc aagtggacaa aataaagggt
      601 gccggtggtg atatcgaagg taggaccatg ggtgatgatg atggtgatga tgatggtgat
      661 gatgattaag gatccgttca actagcagac cattatcaac aaaatactcc aattggcgat
      721 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgccct ttcgaaagat
      781 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
      841 catggcatgg atgagctcta caaactcgag caccaccacc accaccactg agatccggct
      901 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      961 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     1021 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1081 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1141 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1201 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1261 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1321 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1381 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1441 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     1501 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1561 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1621 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1681 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1741 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1801 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1861 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1921 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1981 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2041 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2101 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2161 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2221 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2281 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2341 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2401 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2461 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2521 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2761 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3301 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3361 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3421 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     3481 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3541 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3601 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3661 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3721 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3781 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3841 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3901 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3961 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     4021 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     4081 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     4141 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     4201 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     4261 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     4321 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     4381 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     4441 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     4501 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4561 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4621 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     4681 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4741 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     4801 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     4861 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     4921 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     4981 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     5041 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     5101 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     5161 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     5221 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     5281 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     5341 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     5401 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     5461 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     5521 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     5581 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     5641 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     5701 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     5761 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     5821 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     5881 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     5941 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     6001 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     6061 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     6121 gatctcgatc ccgcgaaatt aatacgactc actatagg
//
