LOCUS       sequence-140285-        6207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses beta-tubulin labeled with Halo-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  AUTHORS   Uno SN, Kamiya M, Yoshihara T, Sugawara K, Okabe K, Tarhan MC, 
            Fujita H, Funatsu T, Okada Y, Tobita S, Urano Y
  TITLE     A spontaneously blinking fluorophore based on intramolecular 
            spirocyclization for live-cell super-resolution imaging.
  JOURNAL   Nat Chem. 2014 Aug;6(8):681-9. doi: 10.1038/nchem.2002. Epub 2014 
            Jul 20.
  PUBMED    25054937
REFERENCE   2  (bases 1 to 6207)
  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: 
            "10.1038/nchem.2002"; journalName: "Nat Chem"; date: "2014-08"; 
            volume: "6"; issue: "8"; pages: "681-9"
COMMENT     Sequence Label: TUBB5-Halo
FEATURES             Location/Qualifiers
     source          1..6207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1993..2877
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    3002..3123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3039..3058)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3093..3112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3130..3585)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3267..3288)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3479..3498
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3612..3716
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3718..4075
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3718..3909
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3741..3761)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3926..4061
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4110..4904
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4110..4904
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4164..4183)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4774..4793
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5092..5111)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5136..5183
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5512..6100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6001..6020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ccaccatgag ggaaatcgtg
      661 cacatccagg ccggacagtg tggcaaccag atcggtgcta agttctggga ggtgataagc
      721 gatgaacatg gcatcgaccc caccggtacc taccacggtg acagcgacct gcagctggac
      781 cgaatctctg tgtactataa tgaagccaca ggtggcaagt atgtccctcg agctatcttg
      841 gtggatctag aacctgggac tatggactcc gttcgctcag gtccttttgg ccagatcttc
      901 agaccagaca acttcgtttt cggtcagtct ggggcaggca acaactgggc taaaggccac
      961 tacacagagg gagctgagtt ggttgactct gtcttggatg tggtgcggaa ggaggcggag
     1021 agctgtgatt gcctgcaagg ctttcagctg acccactcac tgggtggagg cactggctct
     1081 ggcatgggca ccctgctcat cagcaagatc cgggaagaat atcctgaccg tatcatgaat
     1141 accttcagtg tggtgccctc gcccaaagtg tctgataccg tggtcgagcc ctacaatgcc
     1201 accctgtctg tccatcagtt ggttgagaac acggatgaga cctactgcat cgacaacgag
     1261 gccctctacg acatctgctt ccgtaccctc aagctcacca cgccaaccta cggagacctg
     1321 aaccatctcg tctcggccac catgagcggc gtcaccacct gcctccgttt cccgggccag
     1381 cttaatgctg accttcgaaa gctggctgtc aacatggtgc cattcccacg tctccacttc
     1441 ttcatgcctg gctttgcccc tctcaccagc cgtggaagcc agcagtaccg ggccctcact
     1501 gtgcctgaac ttacccagca ggtcttcgat gccaagaaca tgatggccgc ctgcgacccg
     1561 cgccacggcc ggtacctcac agttgccgcc gtcttccgtg gacggatgtc catgaaggag
     1621 gtggatgagc agatgctcaa cgtgcagaac aagaatagca gctacttcgt ggaatggatc
     1681 cccaacaatg tcaagacagc tgtctgtgac atcccaccgc gtggcctcaa gatggcagtc
     1741 accttcattg gaaacagcac agccatccag gagctcttca agcgcatctc tgagcagttt
     1801 acggctatgt tccgccggaa ggctttcctc cactggtata cgggtgaggg catggacgag
     1861 atggagttca ccgaagctga gagcaacatg aacgacctcg tctctgagta ccagcagtac
     1921 caggatgcca ccgcggaaga ggaagaggat ttcggagagg aggcagaaga ggaggcccag
     1981 tcgaccggca gcgaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg
     2041 ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg
     2101 cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc
     2161 catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt
     2221 tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa
     2281 gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc
     2341 aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg
     2401 gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc
     2461 cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat gggtgtcgtc
     2521 cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa tcctgttgac
     2581 cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc
     2641 gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc gaagctgctg
     2701 ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct ggccaaaagc
     2761 ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca agaagacaac
     2821 ccggacctga tcggcagcga gatcgcgcgc tggctgtcca cgctcgagat ttccggctaa
     2881 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2941 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3001 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3061 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3121 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3181 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3241 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3301 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3361 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3421 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3481 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3541 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3601 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3661 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3721 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3781 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3841 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3901 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3961 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4021 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4081 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4141 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4201 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4261 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4321 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4381 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4441 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4501 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4561 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4621 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4681 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4741 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4801 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4861 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4921 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4981 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5041 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5101 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5161 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5221 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5281 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5341 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5401 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5461 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5521 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5581 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5641 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5701 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5761 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5821 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5881 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5941 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6001 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6061 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6121 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6181 tgattctgtg gataaccgta ttaccgc
//
