LOCUS       sequence-204220-        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 K40A
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      605..614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(611..613,614..616,617..1327)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(611..613,614..616,617..1327)
                     /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(656..677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(917..936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1264..1285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2831..2952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2868..2887)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2922..2941
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2959..3414)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3096..3117)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3308..3327
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3441..3545
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3547..3904
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3547..3738
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3570..3590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3755..3890
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3939..4733
                     /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             3939..4733
                     /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(3993..4012)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4603..4622
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4921..4940)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4965..5012
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5341..5929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5830..5849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      601 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      661 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      721 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      781 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      841 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      901 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      961 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1021 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1081 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1141 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1201 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1261 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1321 gtacaagtcc ggactcagat ctcgagtgcg tgagtgcatc tccatccacg ttggccaggc
     1381 tggtgtccag attggcaatg cctgctggga gctctactgc ctggaacacg gcatccagcc
     1441 cgatggccag atgccaagtg acgcgaccat tgggggagga gatgactcct tcaacacctt
     1501 cttcagtgag acgggcgctg gcaagcacgt gccccgggct gtgtttgtag acttggaacc
     1561 cacagtcatt gatgaagttc gcactggcac ctaccgccag ctcttccacc ctgagcagct
     1621 catcacaggc aaggaagatg ctgccaataa ctatgcccga gggcactaca ccattggcaa
     1681 ggagatcatt gaccttgtgt tggaccgaat tcgcaagctg gctgaccagt gcaccggtct
     1741 tcagggcttc ttggttttcc acagctttgg tgggggaact ggttctgggt tcacctccct
     1801 gctcatggaa cgtctctcag ttgattatgg caagaagtcc aagctggagt tctccattta
     1861 cccagcaccc caggtttcca cagctgtagt tgagccctac aactccatcc tcaccaccca
     1921 caccaccctg gagcactctg attgtgcctt catggtagac aatgaggcca tctatgacat
     1981 ctgtcgtaga aacctcgata tcgagcgccc aacctacact aaccttaacc gccttattag
     2041 ccagattgtg tcctccatca ctgcttccct gagatttgat ggagccctga atgttgacct
     2101 gacagaattc cagaccaacc tggtgcccta cccccgcatc cacttccctc tggccacata
     2161 tgcccctgtc atctctgctg agaaagccta ccatgaacag ctttctgtag cagagatcac
     2221 caatgcttgc tttgagccag ccaaccagat ggtgaaatgt gaccctcgcc atggtaaata
     2281 catggcttgc tgcctgttgt accgtggtga cgtggttccc aaagatgtca atgctgccat
     2341 tgccaccatc aaaaccaagc gcagcatcca gtttgtggat tggtgcccca ctggcttcaa
     2401 ggttggcatc aactaccagc ctcccactgt ggtgcctggt ggagacctgg ccaaggtaca
     2461 gagagctgtg tgcatgctga gcaacaccac agccattgct gaggcctggg ctcgcctgga
     2521 ccacaagttt gacctgatgt atgccaagcg tgcctttgtt cactggtacg tgggtgaggg
     2581 gatggaggaa ggcgagtttt cagaggcccg tgaagatatg gctgcccttg agaaggatta
     2641 tgaggaggtt ggtgtggatt ctgttgaagg agagggtgag gaagaaggag aggaatacta
     2701 aggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     2761 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2821 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2881 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2941 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3001 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3061 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3121 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3181 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3241 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3301 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3361 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3421 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3481 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3541 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3601 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3661 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3721 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3781 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3841 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3901 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3961 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4021 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4081 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4141 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4201 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4261 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4321 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4381 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4441 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4501 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4561 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4621 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4681 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4741 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4801 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4861 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4921 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4981 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5041 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5101 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5161 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5221 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5281 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5341 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5401 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5461 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5521 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5581 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5641 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5701 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5761 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5821 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5881 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5941 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6001 gttatcccct gattctgtgg ataaccgtat taccgccatg catta
//
