LOCUS       sequence-345295-        9168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Live-cell marker for stable microtubules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9168)
  AUTHORS   Jansen KI, Iwanski MK, Burute M, Kapitein LC
  TITLE     A live-cell marker to visualize the dynamics of stable microtubules 
            throughout the cell cycle.
  JOURNAL   J Cell Biol. 2023 May 1;222(5):e202106105. doi: 
            10.1083/jcb.202106105. Epub 2023 Mar 2.
  PUBMED    36880745
REFERENCE   2  (bases 1 to 9168)
  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: "J Cell 
            Biol."; date: "2023-05-1"; volume: "222(5):e202106105. doi"; pages: 
            " 10.1083/jcb.202106105. Epub 2023 Mar 2"
COMMENT     Sequence Label: pB80-hKif5b(1-560)rigor-2xmNeongreen
FEATURES             Location/Qualifiers
     source          1..9168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1920..1937)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1920..1936)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1929..1951)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2145..2164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2264..2286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2324..2342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2410..2514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2515..2583,2584..3375)
                     /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(2733..2752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3546..4134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4035..4054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4288..4305
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4422..4443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4458..4463,4464..4481,4482..4488)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4496..4512
                     /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     4501..4523
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4520..4536
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4520..4536
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4550..4684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4574..4593)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4628..4647
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4840..5550)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      5286..5295
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5578..6285)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      6021..6030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      7992..8001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(8070..8092)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cccccccccc cccgccccca tcgctgcaca aaataattaa aaaataaata aatacaaaat
       61 tgggggtggg gagggggggg agatggggag agtgaagcag aacgtggggc tcacctcgag
      121 gtgagggtgt ggggcggggg tgtgacgcgc ccctccatcc cccctccccc ctttttgatt
      181 ccacagtgct ttgttgggga gggggagcgg ggtccgcgtg tgcagccctc cccccacccc
      241 cgaggtggtc ctgcggtccc cttgaagtca cctcccgggg agcgccagca ggagaggcag
      301 ggatagcgat gggcaatgag cgtgcatgca ctgagctgca gcgacactga tgtgcatgca
      361 cgcaccgagc agcaataaca ccggtgtgcg tgcagccact gagctgcagt gatacaatga
      421 gtgtgcgtgc accgagcagc aatcgctcca gtgtgagtgc atgcactggg atgaagtgat
      481 acaatgagcg tgcgtgcaca gagcagcaat aacaccgagg ggtatgcatg cactagactg
      541 ctacacacca aggaggatgc atgcacagcg ctgcaccatc aagacgcatg catgcaccag
      601 gctgcaataa caccaggagc atgcatgcac gggcctggtg ctggggtgca gcctgctggg
      661 atgcaaggga ggggaagggg caccgggtgc aacgcgcgct gcactctcta cacccacccc
      721 gcccggcctg cagcactacc cagggccaag ttcaatgcta atgtcagcac gtagcggtgc
      781 aaacctccag gccggccctt ggactgctga gtgcactgct gggtgcacct cagagcccgt
      841 taagggccac tctttgtcag ggtgtgcgct gcggcggtgg cagccggagg gaggaagcga
      901 gcgctgagta aggagctgag tcaggcaggg cccggcccgg cggaggtcaa ggcagccgaa
      961 tgcctttgcc tacatttggg caatattttg tttctttcct gtttgcagcg cgtgctgcag
     1021 gctgggccgg ggcggtgcgc ggtgttggtg agcagcgctg cgaggggctt tgcagctcca
     1081 gagcggtgca tgagcgcatg ctgattaact gcattgatta aggggccatg caagaagcac
     1141 tgtagatatt aggggagcgg gacccagccc tgtgtgcggt cttgcagtgg ggtctttgtc
     1201 cctgggatgc tgcgaggatg ggttgcatcg gggctgcatt gtgtcccctc caaactggag
     1261 gtggccctgt ggacagagct gagctgcagg gacaccgcag ggacaatact gtctcctcgg
     1321 aggcggtgac actgcagcac atggggacct ccgtgggcac ccagtgccag gaccccgtgg
     1381 gctccatagt gccacatctc tctgtatcag ccccactcgc agcaaaaagg gagtcacact
     1441 gttaattacc tgcctgctgg ggaattaatt aacgtttgtg ctccactttg acaaggccgg
     1501 gtattaatta gaagattata cgtcaaagaa gggcagccac gaggcatggg tacagcggga
     1561 ggtgtgcaga gctgataatg gggcggcaat gggaaatgcc ccgcggtagc catgaagccg
     1621 tgggtttggg ctggggaatg tcaccagaag gcagcgtggg gtgtgagggc ccggcaggat
     1681 gggggtggtg ggggcaatga tggggcaatt cgggttccct tgattatggg gttggatcag
     1741 agcgcggcat ccggagcgtg gcagtgccaa ccccactgtg tgctgctggt agggggctcc
     1801 tcacagtgcc actgccatgc cacagagcca ggaccatcag tttccaaggg gaaccgggtg
     1861 ctgaagggga gcatccccat cctgagagcc gtgggccaat gcaaagatcc caacttggca
     1921 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     1981 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2041 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     2101 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     2161 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2221 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2281 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     2341 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     2401 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2461 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2521 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2581 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2641 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2701 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2761 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2821 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2881 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2941 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3001 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3061 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3121 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3181 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3241 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3301 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3361 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3421 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3481 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3541 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3601 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3661 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3721 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3781 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3841 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3901 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3961 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4021 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4081 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4141 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4201 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4261 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4321 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     4381 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4441 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     4501 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattg atccagacat
     4561 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4621 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4681 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4741 tttttaaagc aagtaaaacc tctacagatg tgatatggct gattatgatc attacttatc
     4801 taggaccggt acgcgttcaa ctagtcatat ggcggccgct tacttgtaca gctcgtccat
     4861 gcccatcaca tcggtaaagg ccttttgcca ctccttgaag ttgagctcgg tcttggagtg
     4921 cttgagctcc gtcttacgga acacgtacat cggctggttc ttcagatagt tagccgccat
     4981 tggcttggca aaggtgtagg tggtccgcgc agtgctccgg tagcgcttgc catttccagt
     5041 ggtgtaactc cacttaaagg tactgatgat ggttttgtcg ttggggtaag tcttcttcga
     5101 cctgcaccag tccgcagcgg tcagcgagtt ggtcatcaca ggaccgtcag cagggaaacc
     5161 agtccccttc acctgggcct ctcctttgat gtggcttccc tcgtaggtgt agcggtagtt
     5221 aacagtaagg gaggcaccat cttcaaactg cattgtgcga tggacttggt agccggagcc
     5281 atctaccatg gcggcctgga aaggcgacat cccgtcaggg tagggcaggt actgatggaa
     5341 gccatacccg atatgaggga ccagaatcca gggggagaac tggaggtcac ccttggtgga
     5401 cttcaggttt aactcctcat aaccatcatt tggattgccg gtgccctgac ccaccatgtc
     5461 aaagtccaca ccgttgatgg agccaaagat gtgtaactca tgtgtcgctg ggagagaggc
     5521 catgttatcc tcctcgccct tgctcaccat actagttgag ccaccgcctc ctctagactt
     5581 gtacagctcg tccatgccca tcacatcggt aaaggccttt tgccactcct tgaagttgag
     5641 ctcggtcttg gagtgcttga gctccgtctt acggaacacg tacatcggct ggttcttcag
     5701 atagttagcc gccattggct tggcaaaggt gtaggtggtc cgcgcagtgc tccggtagcg
     5761 cttgccattt ccagtggtgt aactccactt aaaggtactg atgatggttt tgtcgttggg
     5821 gtaagtcttc ttcgacctgc accagtccgc agcggtcagc gagttggtca tcacaggacc
     5881 gtcagcaggg aaaccagtcc ccttcacctg ggcctctcct ttgatgtggc ttccctcgta
     5941 ggtgtagcgg tagttaacag taagggaggc accatcttca aactgcattg tgcgatggac
     6001 ttggtagccg gagccatcta ccatggcggc ctggaaaggc gacatcccgt cagggtaggg
     6061 caggtactga tggaagccat acccgatatg agggaccaga atccaggggg agaactggag
     6121 gtcacccttg gtggacttca ggtttaactc ctcataacca tcatttggat tgccggtgcc
     6181 ctgacccacc atgtcaaagt ccacaccgtt gatggagcca aagatgtgta actcatgtgt
     6241 cgctgggaga gaggccatgt tatcctcctc gcccttgctc accattaagc ttgccgcttc
     6301 ctggatcgtc gactttttta gtaaagatgc catcatctca gctgctcgtt ttttctggtg
     6361 gttggtcatt tccttaagtt tctgaagctc agcatctata ctcgctaaag ttgccgattt
     6421 ctgattcaat tcatcactaa gcaattcata ttccttagtt ttgtcttcaa cttcctgaga
     6481 cttctgatca taattgacag caagttcttc tagggcctgt aaaacttctt tcacttcttc
     6541 tttagaggca tcattttctg cttgaaggcg attcagctca gcttgcatat tgtcttgatc
     6601 ccttctggta gatgccaaaa gctcctcctg atccaacatt tgcgtcttca gtttctctac
     6661 cagttgactt tgctggttaa tttcttcatc cttgtcatca agctgtttgt ataatttagc
     6721 aatttcttct tcacactttc ttctttcagc atcagtaaaa tttcctataa ctccaattgc
     6781 ggttgctggt ttatcattgg taagagtaat atctttatcc actgtgaaag cttccaagtt
     6841 ggctttctct ttgtcaaact gttcatcaat aggcaccgtc tccccattac gccatctgtt
     6901 gagctcattt tcaagccact gaatagtgtt ccgcaggatc ttattttttt ctttttcttt
     6961 ttcatacttc tttttccact gttctgcagt taactccaca ttgacacaaa ctgtgttctt
     7021 aattgttttg gccctttggc caaataagag tgtagatttt gtttcagact cattgtatga
     7081 tgatggagag cagcaaatta caatagtggt tctacagttg ccacctaatg aatcttgaag
     7141 gattcttgtc attttactat ctcgatatgg aacatatgta ctaccctcag ccaaagcaga
     7201 aataacattt ccaagagcag aaagtgactt gttgatgttt ttagcttcat ccagcacagc
     7261 accttcagct ccagttttac taaccttttc actagcagct aaatcaacca gataaagttt
     7321 tccactcagc ttttgttccg tttgtgtgtt ctcttgtttg acattaataa gaaatatact
     7381 gtgactccta gagctatgtt cattcatatt tgtaactgct acatgtctgt tggattttcc
     7441 ttcatctatg gtatccataa cttcatctgg actacataca aaacgctctg tgcacccctt
     7501 tacataggga actcggtttt tgtcttcatg aactgaaagg ttggtctttg aaacatctaa
     7561 caggtccctt atcttatcca aatatatttc aaaatatgaa accttaatat gaaattccaa
     7621 attttcatcc atggagtaaa tataattaaa aatatcttgc actattcttg gaataattcc
     7681 catgccttct ggatcatgaa gtttaccctc cattgtgtgt gtcttcccag aggatgtttg
     7741 tccatatgca aatattgttc cattatatcc ttcaagtaca tctttaacaa tcttctttgc
     7801 acagtcatta tacacttgct cttgagatgt gcttgactgg aacacccgat caaatgcata
     7861 aggcttggac gcgatcacga ccgtgtcttc tccctgaaac ttggcgatgt acttgtcgcc
     7921 gcggttcact tcagactcgt tgagaggtct gaagcgacac atcactttga tgttgcactc
     7981 ggccaggtcc gccatggtgg cactggcgcg cccaattgcc tagcttgggg gctgcggagg
     8041 aacagagaag ggaagatata aaccccgccg gtcacacgcc agaagccgaa ccccgccctg
     8101 ccccgtcccc cccgaaggca gccgtccccc tgcggacagc cccgaggctg gagatggaga
     8161 aggggacggc ggcgcggcga cgcacgaagg ccctccccgc ccatttcctt cctgccggcg
     8221 ccgcaccgct tcgccccgcg cccgctagag ggggtgcggc ggcgcctccc agatttcggc
     8281 tccgcacaga tttgggacaa aggaagtccc tgcgccctct cgcacgatta ccataaaagg
     8341 caatggctgc ggctcgccgc gcctcgacag ccgccggcgc tccgggggcc gccgcgcccc
     8401 tcccccgagc cctccccggc ccgaggcggc cccgccccgc ccggcacccc cacctgccgc
     8461 caccccccgc ccggcacggc gagccccgcg ccacgccccg cacggagccc cgcacccgaa
     8521 gccgggccgt gctcagcaac tcggggaggg gggtgcaggg gggggttaca gcccgaccgc
     8581 cgcgcccaca ccccctgctc acccccccac gcacacaccc cgcacgcagc ctttgttccc
     8641 ctcgcagccc cccccgcacc gcggggcacc gcccccggcc gcgctcccct cgcgcacact
     8701 gcggagcgca caaagccccg cgccgcgccc gcagcgctca cagccgccgg gcagcgcgga
     8761 gccgcacgcg gcgctcccca cgcacacaca cacgcacgca ccccccgagc cgctcccccc
     8821 cgcacaaagg gccctcccgg agccctttaa ggctttcacg cagccacaga aaagaaacga
     8881 gccgtcatta aaccaagcgc taattacagc ccggaggaga agggccgtcc cgcccgctca
     8941 cctgtgggag taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg gcggcggagc
     9001 ggggcacggg gcgaaggcag cgcgcagcga ctcccgcccg ccgcgcgctt cgctttttat
     9061 agggccgccg ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg ccgctctgat
     9121 tggctgccgc cgcacctctc cgcctcgccc cgccccgccc ckcgcccc
//
