LOCUS       sequence-364435-        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 399, Emission:
            456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  AUTHORS   Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV
  TITLE     An enhanced monomeric blue fluorescent protein with the high 
            chemical stability of the chromophore.
  JOURNAL   PLoS One. 2011;6(12):e28674. Epub 2011 Dec 8.
  PUBMED    22174863
REFERENCE   2  (bases 1 to 5423)
  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: "PLoS One.";
            date: "2011"; volume: "6(12)"; pages: "e28674. Epub 2011 Dec 8"
COMMENT     Sequence Label: mTagBFP2-pBAD
FEATURES             Location/Qualifiers
     source          1..5423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(125..142)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      295..381
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      473..500
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        519..610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(611..679,680..1471)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(829..848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1513..1968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1600..1619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1810..1831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2079..2667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2568..2587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2821..2838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2853..2993
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2919..2938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3079..3101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3239..3258
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3438..4316)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4343..4627
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4550..4569
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4654..4676
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4696..4713
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4717..4749
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             4762..4782
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4789..5423,1..76)
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
ORIGIN
        1 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact
       61 ggggcacaag cttaattaag aattcgaagc ttggctgttt tggcggatga gagaagattt
      121 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
      181 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
      241 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
      301 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      361 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
      421 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
      481 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
      541 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      601 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      661 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      721 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      781 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      841 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      901 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      961 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1021 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1081 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1141 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1201 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1261 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1321 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1381 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1441 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1501 ttagattgat ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1561 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1621 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1681 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     1741 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1801 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     1861 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1921 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttaa aaggatctag
     1981 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2041 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2221 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2641 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2701 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2761 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2821 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     2881 ctgtgcggta tttcacaccg catctggtgc actctcagta caatctgctc tgatgccgca
     2941 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3001 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3061 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3121 aacgcgcgag gcagaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     3181 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     3241 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     3301 tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc
     3361 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt
     3421 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa
     3481 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag
     3541 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc
     3601 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct
     3661 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg
     3721 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt
     3781 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac
     3841 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg
     3901 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag
     3961 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga
     4021 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga
     4081 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga
     4141 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc
     4201 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc
     4261 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt
     4321 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt
     4381 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc
     4441 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat
     4501 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca
     4561 tttttatcca taagattagc ggatactacc tgacgctttt tatcgcaact ctctactgtt
     4621 tctccatacc cgtttttttg ggctagaaat aattttgttt aactttaaga aggagatata
     4681 catatgcggg gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag
     4741 caaatgggtc gggatctgta cgagaacctg tacttccagg gctcgagcat ggtgtctaag
     4801 ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     4861 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     4921 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     4981 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     5041 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     5101 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     5161 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     5221 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     5281 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     5341 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     5401 agaatcaagg aggccaacaa cga
//
