LOCUS       sequence-271345-        5408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 588, Emission:
            635. Also known as mKate2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5408)
  AUTHORS   Shcherbo D, Murphy CS, Ermakova GV, Solovieva EA, Chepurnykh TV, 
            Shcheglov AS, Verkhusha VV, Pletnev VZ, Hazelwood KL, Roche PM, 
            Lukyanov S, Zaraisky AG, Davidson MW, Chudakov DM
  TITLE     Far-red fluorescent tags for protein imaging in living tissues.
  JOURNAL   Biochem J. 2009 Mar 15;418(3):567-74. doi: 10.1042/BJ20081949.
  PUBMED    19143658
REFERENCE   2  (bases 1 to 5408)
  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"; journalName: 
            "Biochem J"; date: "2009-03-15- 15"; volume: "418"; issue: "3"; 
            pages: "567-74"
COMMENT     Sequence Label: mKate1.31-pBAD
FEATURES             Location/Qualifiers
     source          1..5408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    148..288
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(214..233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     374..396
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     534..553
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(733..1611)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1638..1922
                     /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     1845..1864
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1949..1971
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1991..2008
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2012..2044
                     /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             2057..2077
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2084..2782
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     primer_bind     complement(2828..2845)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2998..3084
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3176..3203
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3222..3313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3314..3382,3383..4174)
                     /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(3532..3551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4216..4671
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4303..4322)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4513..4534
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4782..5370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5271..5290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
       61 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
      121 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
      181 cggtatttca caccgcatct ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
      241 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
      301 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
      361 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
      421 gcgaggcaga aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
      481 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
      541 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
      601 ccggcgtaga ggatctgctc atgtttgaca gcttatcatc gatgcataat gtgcctgtca
      661 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
      721 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
      781 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
      841 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
      901 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
      961 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     1021 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     1081 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     1141 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     1201 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     1261 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     1321 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     1381 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     1441 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     1501 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     1561 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     1621 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     1681 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     1741 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     1801 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     1861 atccataaga ttagcggata ctacctgacg ctttttatcg caactctcta ctgtttctcc
     1921 atacccgttt ttttgggcta gaaataattt tgtttaactt taagaaggag atatacatat
     1981 gcggggttct catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat
     2041 gggtcgggat ctgtacgaga acctgtactt ccagggctcg agcatggtga gcgagctgat
     2101 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     2161 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggc
     2221 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     2281 cagcaaaacc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
     2341 cgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     2401 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     2461 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg cctccaccga
     2521 gaccctgtac cccgctgacg gcggcctgga aggcagagcc gacatggccc tgaagctcgt
     2581 gggcgggggc cacctgatct gcaacttgaa gaccacatac agatccaaga aacccgctaa
     2641 gaacctcaag atgcccggcg tctactatgt ggacagaaga ctggaaagaa tcaaggaggc
     2701 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     2761 tagcaaactg gggcacagat gagaattcga agcttggctg ttttggcgga tgagagaaga
     2821 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     2881 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     2941 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     3001 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3061 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     3121 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     3181 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     3241 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3301 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3361 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3421 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3481 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3541 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     3601 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3661 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3721 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3781 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3841 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3901 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3961 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4021 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4081 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4141 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4201 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4261 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4321 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4381 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4441 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4501 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     4561 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4621 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc
     4681 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4741 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4801 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4861 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4921 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4981 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5041 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5101 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5161 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5221 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5281 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5341 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5401 ctggcctt
//
