LOCUS       sequence-125710-        3760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the CDS of the Blue fluorescente protein as a level 0 
            GoldenBraid part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3760)
  AUTHORS   Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, 
            Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D
  TITLE     GoldenBraid 2.0: a comprehensive DNA assembly framework for plant 
            synthetic biology.
  JOURNAL   Plant Physiol. 2013 Jul;162(3):1618-31.
  PUBMED    23669743
REFERENCE   2  (bases 1 to 3760)
  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: "Plant 
            Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31"
COMMENT     Sequence Label: pBFP (GB0025)
FEATURES             Location/Qualifiers
     source          1..3760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..707
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        complement(810..828)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(811..828)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(846..862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(846..862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(859..881)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    870..886
                     /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)."
     promoter        complement(join(894..900,901..918,919..924))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    939..960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1077..1094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1248..1836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1328..1347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2007..2798,2799..2867))
                     /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     2630..2649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2868..2972)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3050..3505)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3187..3208)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3399..3418
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3631..3653
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3645..3662
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3646..3662
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3669..3688
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3669..3687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3718..3727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3722..3731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcgaatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
       61 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
      121 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
      181 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
      241 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
      301 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
      361 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
      421 gctgggaggc cttcaccgaa acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
      481 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag accacatata
      541 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
      601 tggaaagaat caaggaggcc aacaacgaaa cctacgtcga gcagcacgag gtggcagtgg
      661 ccagatactg cgacctccct agcaaactgg ggcacaagct taattgagct tcgagaccac
      721 tcatcgccca atcactagtg aattcgcggc cgcctgcagg tcgaccatat gggagagctc
      781 ccaacgcgtt ggatgcatag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa
      841 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      901 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      961 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1021 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1081 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1141 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1201 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1261 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1321 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1381 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1441 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1501 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1561 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1621 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1681 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1741 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1801 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1861 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1921 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1981 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2041 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2101 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2161 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2221 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2281 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2341 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2401 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2461 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2521 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2581 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2641 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2701 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2761 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2821 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2881 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2941 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat
     3001 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     3061 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3121 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3181 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3241 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3301 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3361 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3421 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3481 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     3541 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3601 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3661 gtgaattgta atacgactca ctatagggcg aattgggccc gacgtcgcat gctcccggcc
     3721 gccatggcgg ccgcgggaat tcgattgggc gatgagtggt
//
