LOCUS       sequence-377383-        7356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-tagged MASTL WT with resistance to MASTL siRNA 
            (ACGCCTTATTCTAGCAAATTA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7356)
  AUTHORS   Taskinen ME, Narva E, Conway JRW, Hinojosa LS, Lilla S, Mai A, De 
            Franceschi N, Elo LL, Grosse R, Zanivan S, Norman JC, Ivaska J
  TITLE     MASTL promotes cell contractility and motility through 
            kinase-independent signaling.
  JOURNAL   J Cell Biol. 2020 Jun 1;219(6). pii: 151688. doi: 
            10.1083/jcb.201906204.
  PUBMED    32311005
REFERENCE   2  (bases 1 to 7356)
  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: "2020-06-1"; volume: "219(6). pii: 151688. doi"; 
            pages: " 10.1083/jcb.201906204"
COMMENT     Sequence Label: pcDNA-EGFP MASTL WT (siRNA resistant)
FEATURES             Location/Qualifiers
     source          1..7356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(632..634,635..637,638..1348)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(632..634,635..637,638..1348)
                     /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(677..698)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(938..957)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1285..1306
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4163..4284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4200..4219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4254..4273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4291..4746)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4640..4659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4773..4877
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4879..5236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4879..5070
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4902..4922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5087..5222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5271..6065
                     /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             5271..6065
                     /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(5325..5344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5935..5954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6253..6272)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6297..6344
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6673..7261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7162..7181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      601 gtcagatccg ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
      661 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      721 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      781 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      841 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      901 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      961 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1021 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1081 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1141 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1201 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     1261 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1321 cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttccat
     1381 ggatcccacc gcgggaagca agaaggagcc tggaggaggc gcggcgactg aggagggcgt
     1441 gaataggatc gcagtgccaa aaccgccctc cattgaggaa ttcagcatag tgaagcccat
     1501 tagccggggc gccttcggga aagtgtatct ggggcagaaa ggcggcaaat tgtatgcagt
     1561 aaaggttgtt aaaaaagcag acatgatcaa caaaaatatg actcatcagg tccaagctga
     1621 gagagatgca ctggcactaa gcaaaagccc attcattgtc catttgtatt attcactgca
     1681 gtctgcaaac aatgtctact tggtaatgga atatcttatt gggggagatg tcaagtctct
     1741 cctacatata tatggttatt ttgatgaaga gatggctgtg aaatatattt ctgaagtagc
     1801 actggctcta gactaccttc acagacatgg aatcatccac agggacttga aaccggacaa
     1861 tatgcttatt tctaatgagg gtcatattaa actgacggat tttggccttt caaaagttac
     1921 tttgaataga gatattaata tgatggatat ccttacaaca ccatcaatgg caaaacctag
     1981 acaagattat tcaagaaccc caggacaagt gttatcgctt atcagctcgt tgggatttaa
     2041 cacaccaatt gcagaaaaaa atcaagaccc tgcaaacatc ctttcagcct gtctgtctga
     2101 aacatcacag ctttctcaag gactcgtatg ccctatgtct gtagatcaaa aggacactac
     2161 cccctacagc tctaagctgc taaaatcatg tcttgaaaca gttgcctcca acccaggaat
     2221 gcctgtgaag tgtctaactt ctaatttact ccagtctagg aaaaggctgg ccacatccag
     2281 tgccagtagt caatcccaca ccttcatatc cagtgtggaa tcagaatgcc acagcagtcc
     2341 caaatgggaa aaagattgcc aggaaagtga tgaagcattg ggcccaacaa tgatgagttg
     2401 gaatgcagtt gaaaagttat gcgcaaaatc tgcaaatgcc attgagacga aaggtttcaa
     2461 taaaaaggat ctggagttag ctctttctcc cattcataac agcagtgccc ttcccaccac
     2521 tggacgctct tgtgtaaacc ttgctaaaaa atgcttctct ggggaagttt cttgggaagc
     2581 agtagaactg gatgtaaata atataaatat ggacactgac acaagtcagt taggtttcca
     2641 tcagtcaaat cagtggactg tggattctgg tgggatatct gaagagcacc ttgggaaaag
     2701 aagtttaaaa agaaattttg agttggttga ctccagtcct tgtaaaaaaa ttatacagaa
     2761 taaaaaaact tgtgtagagt ataagcataa cgaaatgaca aattgttata caaatcaaaa
     2821 tacaggctta acagttgaag tgcaggacct taagctatca gtgcacaaaa gtcaacaaaa
     2881 tgactgtgct aataaggaga acattgtcaa ttcttttact gataaacaac aaacaccaga
     2941 aaaattacct ataccaatga tagcaaaaaa ccttatgtgt gaactcgatg aagactgtga
     3001 aaagaatagt aagagggact acttaagttc tagttttcta tgttctgatg atgatagagc
     3061 ttctaaaaat atttctatga actctgattc atcttttcct ggaatttcta taatggaaag
     3121 tccattagaa agtcagccct tagattcaga tagaagcatc aaagaatcct cttttgaaga
     3181 atcaaatatt gaagatccac ttattgtaac accagattgc caagaaaaga cctcaccaaa
     3241 aggtgtcgag aaccctgctg tacaagagag taaccaaaaa atgttaggtc ctcctttgga
     3301 ggtgctgaaa acgttagcct ctaaaagaaa tgctgttgct tttcgaagtt ttaacagtca
     3361 tattaatgca tccaataact cagaaccatc cagaatgaac atgacttctt tagatgcaat
     3421 ggatatttcg tgtgcctaca gtggttcata tcccatggct ataaccccta ctcaaaaaag
     3481 aagatcctgt atgccacatc agcagacccc aaatcagatc aagtcgggaa ctccataccg
     3541 aactccgaag agtgtgagaa gaggggtggc ccccgttgat gatgggcgaa ttctaggaac
     3601 cccagactac cttgcacctg agctgttact aggcagggcc catggtcctg cggtagactg
     3661 gtgggcactt ggagtttgct tgtttgaatt tctaacagga attccccctt tcaatgatga
     3721 aacaccacaa caagtattcc agaatattct gaaaagagat atcccttggc cagaaggtga
     3781 agaaaagtta tctgataatg ctcaaagtgc agtagaaata cttttaacca ttgatgatac
     3841 aaagagagct ggaatgaaag agctaaaacg tcatcctctc ttcagtgatg tggactggga
     3901 aaatctgcag catcagacta tgcctttcat cccccagcca gatgatgaaa cagatacctc
     3961 ctattttgaa gccaggaata ctgctcagca cctgaccgta tctggattta gtctgtaggg
     4021 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     4081 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     4141 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     4201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4261 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     4321 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4381 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4441 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4501 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4561 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4621 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4681 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4741 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4801 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4861 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4981 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5041 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5101 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5161 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5221 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5281 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5341 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5401 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     5461 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5521 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5581 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5641 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5701 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5761 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5821 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5881 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5941 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6001 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6061 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6121 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6181 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     6241 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     6301 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     6361 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     6421 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     6481 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6541 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6601 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6661 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6721 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6781 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6841 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6901 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6961 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7021 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7081 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7141 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7201 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7261 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7321 gttctttcct gcgttatccc ctgattctgt ggataa
//
