LOCUS       sequence-98502-e        8318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of Multibow set of constructs--each FP gene is initially not 
            expressed and then adopts a permanent ON or OFF status upon 
            Cre-mediated recombination. Encodes cytoplasmic mKate2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8318)
  AUTHORS   Xiong F, Obholzer ND, Noche RR, Megason SG
  TITLE     Multibow: digital spectral barcodes for cell tracing.
  JOURNAL   PLoS One. 2015 May 26;10(5):e0127822. doi: 
            10.1371/journal.pone.0127822. eCollection 2015.
  PUBMED    26010570
REFERENCE   2  (bases 1 to 8318)
  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: "2015-05-26"; volume: "10(5):e0127822. doi"; pages: " 
            10.1371/journal.pone.0127822. eCollection 2015"
COMMENT     Sequence Label: pMTB-Multibow-fR
FEATURES             Location/Qualifiers
     source          1..8318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      353..941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     842..861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1095..1112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1229..1250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1265..1270,1271..1288,1289..1295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1319
                     /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)."
     primer_bind     1308..1330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1327..1343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1327..1343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1362..1382
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1364..1382
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(1459..1470,1471..1658)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        4373..4391
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4373..4390
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(4446..4479)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    complement(4487..4520)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(4530..4563)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4577..4586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5280..5401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(5484..5517)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      5531..5540
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5570..6265
                     /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"
     polyA_signal    6469..6590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6605..6638)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(6692..6829,6830..6841)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6896..6913)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6896..6912)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6905..6927)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7054..7509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7141..7160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7351..7372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7535..7639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7640..7708,7709..8318,1..182)
                     /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     complement(7858..7877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
       61 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      361 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      421 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      481 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
      541 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      601 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      661 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      721 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      781 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      841 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      901 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      961 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1021 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1081 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     1141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1201 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1261 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1321 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     1381 gggaacaaaa gctggagctc caccgcggtg gcggcctagt ggatctgctg ggcttgctga
     1441 aggtaggggg tcaagaacca gaggtgtaaa gtacttgagt aattttactt gattactgta
     1501 cttaagtatt atttttgggg atttttactt tacttgagta caattaaaaa tcaatacttt
     1561 tacttttact taattacatt tttttagaaa aaaaagtact ttttactcct tacaatttta
     1621 tttacagtca aaaagtactt attttttgga gatcacttgg gcccggctcg agggcctagt
     1681 cattacctat actcaacgtt tactctgaga aacttccata ttttagaggt aaaatgctca
     1741 gaattgtaaa aagtcttgtc aaaatacttg gaatttgcca aaaacaaaat aaaagtcaca
     1801 tcacctcatt gtccttggta tcagcaagca cttttaacac aatcagctgt ttttgtttat
     1861 aactcccact gaaggccatg tgcattactc ccaagttaca cagtaaaaag tcacatgtag
     1921 tttatcatga tcactgctgc tcaagcacag ctttatctgt tattcagtgt ggcaatatct
     1981 gaaatgtgtg caaaatatta tttattgtag ctgatcacca acactttcag tgtttttttt
     2041 ttctactttt aatgagaatt atgttcttag gatttatcct tataaacctc tttaaaaaga
     2101 aatgttatgt ctgcatagac acgacatgaa attatgaaca gaaaatttaa ctatttaaga
     2161 tcaagacttg acatataaca tttctaatga acattaaacc tgaggtaggc ccacttgcaa
     2221 aaatgtaact gctatcactc actttataca tcattgtgta acttgtacac acaaaaatag
     2281 tgaagcaatg tggcatggaa tgcaggtccc attcattcaa agttattaaa gcactttctt
     2341 aaagtgcatt actactgagc taagaccagc tagttgctca tacaacatac tacaagtagt
     2401 gtcatcctaa tttggataaa aaaaatgtcc attttataca tggaaaagta ctcattagat
     2461 cccccattgt ttgtattacg gtatttcgtg aacacaagag gtaaatgacc tacagagctg
     2521 ctgttgtgtt agattgaaaa cacaacacag gatcatggag gcatccatca ttatgacgca
     2581 ttctgccact tgtaactctg cacagacttt ttgaaaagtt taattgagtc ttggcgttgt
     2641 cgcagaaaaa tacgaaaaat tctgtgttaa aatttgcaaa taaaaattcc agttttatag
     2701 gaaatcgcca tggccttgtt ctttaaccag ttaagcttcc ccttctttca ctctcaagtt
     2761 gcaagaagca agtgtagcaa tgtgcacgcg acagccgggt gtgtgacgct ggaccaatca
     2821 gagcacagag ctccgaaagt ttacctttta tggctagagc cgggtatgtg ccgtcatata
     2881 aaagagcgcg cccagctttt cagcctcact ttgagctcct ccacacgcag ctagtgcgga
     2941 atatcatctg cttgtaaccc attctcttaa gtcgacaacc ccccaaaccc aaggtgagtt
     3001 ggttttacag tttttaagcg tttaattata gtttctatct attaatgtta agtaaaatgt
     3061 gtaaatggat aaatatgatt tgatgaagtc tttagacagt tcataacggt ctctttgtag
     3121 ttatctacta ggtttgattt ataagtggtg gttcttttga taatctatgt tacatttaat
     3181 aaatattggc tgtatcttaa cttttttgta gcgttagtgg atgtggcagg tgagaatgtc
     3241 ggtaatttaa cgtgaccagt cgtaggcacg acattgaaca ttgaataggc cggtgtgaaa
     3301 tgggtgttaa gtcttaatat aaagttgtgt tctggctaat ctctctttaa cagccataaa
     3361 tgtgattttt aactacagtt ttgagggtgc tggtgagcat gttgcacact tgatggatag
     3421 ccggcatggg aagttctttg tgcaggcagt gctgcagcag ggtgtgacct actttagcta
     3481 gccggctaac cagctctcat ctgatgtaac gtaaacccca tcaatctaac atcggctcat
     3541 tcgtgcttta taagatcgaa gatcttgtta ttagtaacgg gtattggcat ttagttttaa
     3601 cttggttaat tttctttttg ttatgtgttc tatttaaatg tagcctgcag ccccctctgt
     3661 ctttaatgaa gctgaagatt aacgttaacg ttagtgaagt gttgtatccg ctatgaaaga
     3721 tttgaatttt agaggtgtgg tccactttaa taacggctgc ggagttctct ggctgcactt
     3781 cagccgctgt gactgtcgaa gcagagccca tcagtttttc aggatctcgg tgatttttga
     3841 ggctagacta tgaactgaac cgactgagag tgaccgtcca gaaatgcttt agtgtgaagt
     3901 ctccgctcac gaggagtccg tggactgact gcagatctgt gacgcagtaa cacccggcgg
     3961 cagacacccg ttgaaatgcg gttgtgtaat tgatcgcagg cgagggtgga agaggatgtg
     4021 aaacttcatt tgtgtaaaat ttagggagtg gtcctggtgt gatgaatgtc gaaatctgtt
     4081 cctttttact gagccctacg actctggctg agtgccacac cgccggcagc cgcaaagcgg
     4141 ctcaaaccat tgccttttat ggtaataatg agagaatgca gagggacttc ctttgtctgg
     4201 catatctgag gcgcgcgctg tcactagcgc ccaccagcgg tcagactgta gaatgcagag
     4261 caaaacagga agttgactcc agatggtcac atgcttgctg aaacgcctct ccctggcagc
     4321 agtgcaattc taacagtttc cctttctttt acagttcagc catcctaggc ctatttaggt
     4381 gacactatag aagagtacaa gctacttgtt ctttttgcag gatccgccac catgactagc
     4441 tcgagataac ttcgtatagt ataccttata cgaagttatc gagctcataa cttcgtatag
     4501 gatactttat acgaagttat aacgtcgaga taacttcgta tagcatacat tatacgaagt
     4561 tataccggca gatctcgcca ccatggtgag tgtgattaaa ccagagatga agatgaagta
     4621 cttcatggac ggctccgtca atgggcatga gttcacagtt gaaggtgaag gcacaggcaa
     4681 accttacgag ggacatcaag agatgacact acgcgtcaca atggccaagg gcgggccaat
     4741 gcctttctcg tttgacttag tgtcacacac gttctgttac ggccacagac cttttactaa
     4801 atatccagaa gagataccag actatttcaa acaagcattt cctgaaggcc tgtcatggga
     4861 aaggtcgttg cagttcgtgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     4921 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4981 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     5041 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     5101 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaacacgtgc
     5161 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     5221 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     5281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5401 aaggcgtaaa ttgtaagcgt taatgctccg agggccgcca ccacctgttc ctgtaggaat
     5461 tggccgcgac tctaggatct aggataactt cgtataggat actttatacg aagttattga
     5521 tcaattcaca gccaccatgg gagcgccagc tggaggtgca ggtgcagcaa tggtgtctga
     5581 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca
     5641 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     5701 caaggccgtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat
     5761 gtacggcagc aaaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc
     5821 cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
     5881 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     5941 ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctc
     6001 caccgagacc ctgtaccccg ctgacggcgg cctggaaggc agagccgaca tggccctgaa
     6061 gctcgtgggc gggggccacc tgatctgcaa cttgaagacc acatacagat ccaagaaacc
     6121 cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac agaagactgg aaagaatcaa
     6181 ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga
     6241 cctccctagc aaactggggc acagagcagc agcatgataa atggtcctgc tggagttcgt
     6301 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     6361 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     6421 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     6481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgcgatt
     6601 cgagataact tcgtatagca tacattatac gaagttatac cggaattgtc gcgatcgccg
     6661 aattatcccg ggtttaaacg catgcagatc taatactcaa gtacaatttt aatggagtac
     6721 ttttttactt ttactcaagt aagattctag ccagatactt ttacttttaa ttgagtaaaa
     6781 ttttccctaa gtacttgtac tttcacttga gtaaaatttt tgagtacttt ttacacctct
     6841 gtcaagaacc atatgccggt acccaattcg ccctataggc gtattacgcg cgctcactgg
     6901 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     6961 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     7021 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     7081 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     7141 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     7201 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7261 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7321 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7381 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     7441 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     7501 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7561 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7621 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7681 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7741 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7801 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7861 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7921 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7981 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     8041 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     8101 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     8161 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     8221 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     8281 tgcaggacca cttctgcgct cggcccttcc ggctggct
//
