LOCUS       sequence-204218-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To study effects of acetylation-null tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  AUTHORS   Joo EE, Yamada KM
  TITLE     MYPT1 regulates contractility and microtubule acetylation to 
            modulate integrin adhesions and matrix assembly.
  JOURNAL   Nat Commun. 2014 Mar 25;5:3510. doi: 10.1038/ncomms4510.
  PUBMED    24667306
REFERENCE   2  (bases 1 to 6045)
  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: "Nat 
            Commun."; date: "2014-03-25"; volume: "5:3510. doi"; pages: " 
            10.1038/ncomms4510"
COMMENT     Sequence Label: pEGFP Tubulin K40R
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     514..533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        788..1091
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1092..1295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1246..1266
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1334..1343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1340..1342,1343..1345,1346..2056)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1340..1342,1343..1345,1346..2056)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1385..1406)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1646..1665)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1993..2014
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3560..3681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3597..3616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3651..3670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3688..4143)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3825..3846)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4037..4056
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4170..4274
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4276..4633
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4276..4467
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4299..4319)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4484..4619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4668..5462
                     /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             4668..5462
                     /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(4722..4741)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5332..5351
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5650..5669)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5694..5741
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
       61 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      721 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      781 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      841 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      901 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      961 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1021 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1081 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     1141 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     1201 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     1261 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
     1321 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1381 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1441 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1501 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1561 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1621 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1681 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1741 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1801 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1861 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1921 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1981 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2041 ggacgagctg tacaagtccg gactcagatc tcgagtgcgt gagtgcatct ccatccacgt
     2101 tggccaggct ggtgtccaga ttggcaatgc ctgctgggag ctctactgcc tggaacacgg
     2161 catccagccc gatggccaga tgccaagtga caggaccatt gggggaggag atgactcctt
     2221 caacaccttc ttcagtgaga cgggcgctgg caagcacgtg ccccgggctg tgtttgtaga
     2281 cttggaaccc acagtcattg atgaagttcg cactggcacc taccgccagc tcttccaccc
     2341 tgagcagctc atcacaggca aggaagatgc tgccaataac tatgcccgag ggcactacac
     2401 cattggcaag gagatcattg accttgtgtt ggaccgaatt cgcaagctgg ctgaccagtg
     2461 caccggtctt cagggcttct tggttttcca cagctttggt gggggaactg gttctgggtt
     2521 cacctccctg ctcatggaac gtctctcagt tgattatggc aagaagtcca agctggagtt
     2581 ctccatttac ccagcacccc aggtttccac agctgtagtt gagccctaca actccatcct
     2641 caccacccac accaccctgg agcactctga ttgtgccttc atggtagaca atgaggccat
     2701 ctatgacatc tgtcgtagaa acctcgatat cgagcgccca acctacacta accttaaccg
     2761 ccttattagc cagattgtgt cctccatcac tgcttccctg agatttgatg gagccctgaa
     2821 tgttgacctg acagaattcc agaccaacct ggtgccctac ccccgcatcc acttccctct
     2881 ggccacatat gcccctgtca tctctgctga gaaagcctac catgaacagc tttctgtagc
     2941 agagatcacc aatgcttgct ttgagccagc caaccagatg gtgaaatgtg accctcgcca
     3001 tggtaaatac atggcttgct gcctgttgta ccgtggtgac gtggttccca aagatgtcaa
     3061 tgctgccatt gccaccatca aaaccaagcg cagcatccag tttgtggatt ggtgccccac
     3121 tggcttcaag gttggcatca actaccagcc tcccactgtg gtgcctggtg gagacctggc
     3181 caaggtacag agagctgtgt gcatgctgag caacaccaca gccattgctg aggcctgggc
     3241 tcgcctggac cacaagtttg acctgatgta tgccaagcgt gcctttgttc actggtacgt
     3301 gggtgagggg atggaggaag gcgagttttc agaggcccgt gaagatatgg ctgcccttga
     3361 gaaggattat gaggaggttg gtgtggattc tgttgaagga gagggtgagg aagaaggaga
     3421 ggaatactaa ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     3481 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3541 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3601 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3661 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3721 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3781 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3841 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3901 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3961 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4021 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4081 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4141 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4201 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4261 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4321 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4381 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4441 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4501 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4561 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4621 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4681 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4741 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4801 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4861 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4921 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4981 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5041 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5101 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5161 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5221 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5281 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5341 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5401 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5461 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5521 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5581 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5641 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5701 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5761 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5821 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5881 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccc
//
