LOCUS       sequence-244985-        4175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NADPH-dependent blue fluorescent protein .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4175)
  AUTHORS   Hwang CS, Choi ES, Han SS, Kim GJ
  TITLE     Screening of a highly soluble and oxygen-independent blue 
            fluorescent protein from metagenome.
  JOURNAL   Biochem Biophys Res Commun. 2012 Mar 23;419(4):676-81. doi: 
            10.1016/j.bbrc.2012.02.075. Epub 2012 Feb 20.
  PUBMED    22382031
REFERENCE   2  (bases 1 to 4175)
  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: 
            "10.1016/j.bbrc.2012.02.075"; journalName: "Biochem Biophys Res 
            Commun"; date: "2012-03-23- 23"; volume: "419"; issue: "4"; pages: 
            "676-81"
COMMENT     Sequence Label: pQE30-mBFP
FEATURES             Location/Qualifiers
     source          1..4175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      922..1016
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1060..1719
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1126..1145)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1784..1870
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1917..1939)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2025..2165
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2080..2099
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2180..2197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2351..2939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2431..2450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3110..3901,3902..3970))
                     /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     3733..3752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3971..4075)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4143..4161
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc gcatgccaga atctgaacgg caaagtggct
      181 ttcgtgaccg gcggcagccg cggcatcggc gcggcgatcg tccgccgctt ggcggcggac
      241 ggcgccgaca tcgcgttcac ctatgtcagc gcctcgtcga aaaacgtggc caccgccctg
      301 gtgcaagaac tcgaggccaa gggccgccgc gctcgcgcca tccaggcgga ctcggcggat
      361 ccggcccagg tgcggcaggc ggtcgagcag gccatcgtgc aactggggcc ggtggacgtg
      421 ctggtgaaca acgccggcat cttcctggcc ggccccttgg gcgaggtgac gctggacgac
      481 tacgaacgca cgatgaacat caatgtgcgc gcgcctttcg tggccatcca ggccgcgcag
      541 gcctcgatgc cggacggcgg ccgcatcatc aacatcggca gctgcctggc ggaacgcgcc
      601 ggccgagccg gggtaacgct gtatgccgcc agcaagtcgg cgctgctggg catgacgcgc
      661 ggcctggcgc gcgacctggg cgcgcgcggc atcaccgcca acgtcgtgca cccgggcccg
      721 atcgacaccg acatgaatcc cgcagatggc gaacgctcgg gcgaactggt ggccgtgctg
      781 tccttgcctc attacggcga ggtgcgcgac atcgccggca tggtggcttt cctggccggg
      841 ccggatgggc gctacgtgac cggtgcgagt ctggcggtgg acggcggctt cgccgcttga
      901 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
      961 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
     1021 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1081 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1141 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1201 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1261 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1321 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1381 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1441 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1501 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1561 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1621 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1681 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     1741 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     1801 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1861 taggacaaat ccgccctcta gagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     1921 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     1981 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     2041 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     2101 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2161 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2221 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2281 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2341 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2401 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2521 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2761 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2941 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3001 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3061 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3121 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3181 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3241 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3301 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3361 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3421 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3481 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3541 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3601 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3661 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3721 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3781 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3841 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3901 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3961 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4021 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4081 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4141 aaaataggcg tatcacgagg ccctttcgtc ttcac
//
