LOCUS       sequence-271042-        4944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse MyoGEF-T575A mutant in Gateway pENTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4944)
  AUTHORS   de Carcer G, Wachowicz P, Martinez-Martinez S, Oller J, 
            Mendez-Barbero N, Escobar B, Gonzalez-Loyola A, Takaki T, El Bakkali
            A, Camara JA, Jimenez-Borreguero LJ, Bustelo XR, Canamero M, Mulero 
            F, de Los Angeles Sevilla M, Montero MJ, Redondo JM, Malumbres M
  TITLE     Plk1 regulates contraction of postmitotic smooth muscle cells and is
            required for vascular homeostasis.
  JOURNAL   Nat Med. 2017 Aug;23(8):964-974. doi: 10.1038/nm.4364. Epub 2017 Jul
            10.
  PUBMED    28692064
REFERENCE   2  (bases 1 to 4944)
  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/nm.4364"; 
            journalName: "Nat Med"; date: "2017-08"; volume: "23"; issue: "8"; 
            pages: "964-974"
COMMENT     Sequence Label: pENTR-Mm_MyoGEF(T575A)
FEATURES             Location/Qualifiers
     source          1..4944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3069..3168)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3185..3204)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3186..3204)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3209..3225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3209..3225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3327..3346)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3338..4147
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3405..3424)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4294..4882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4783..4802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat gcaagccttt ggtcctccaa atgagggccc
      721 cctccaaggc ctggtagctt cccgaattga gacttacggg ggccgtcatc agacctctgc
      781 ccacagcact gctgggaatc tatttcccag aggaggtcct ggagtggacc ctagtcgccg
      841 acgccttcag cactatgtcc cctttgccaa gggctctggc cagacccgag gcatgtctcc
      901 tctggtgctt cgggaaccag acccagagaa gaggcatgga agctattttg gggtcggccc
      961 acctcactcc cccaaactca aggaagtcac cagggcccat gagctagaaa taaggctgca
     1021 cacattcagc atgtttggga tgccccgcct gccaccagag gaccggagac actgggagat
     1081 aggagaggga ggtgatggtg ccctgaccat ggaaaagtcc tggaaggagc tggtgctgga
     1141 acacaaggag atgaaccgcc agctttgcca ccagcaggaa gcactgtggg aactcctgac
     1201 cactgagctg atctacctga gaaagctcaa gatcatgact gatctcctgg cagctggtct
     1261 gctgaatttg cagcgggtgg ggctgctgac tgaggtgtca gctgagaccc tgtttggaaa
     1321 tgtccccaac ctgattagag cccaccagag attttgggag gaggtgctgc agcccatcct
     1381 ggaagacact cgcacctctg gccaaccttt ggatcctgtc agcttacaaa atggcttcct
     1441 gacatttggc cagcggttcc aaccttacgt ccagtactgc cttcgagtaa agcagaccat
     1501 ggcttatgcg cgggagcagc aagacacgaa cccactgttc cacaccttcg tgcagtggtg
     1561 tgagaaacac aagcgctccg gaagacagac gctgggtgac ttgctcatca agccccacca
     1621 gcgcatcacc aaatacccac tgctgctgca ggctgtgcgc aagagaagcc ccgagccccg
     1681 agcccaagag gccctgaatg ccatgatcga ggctgtggag tccttcctgc gacacataaa
     1741 tggccaggtc cgacagggtg aagagcagga gagtttgatg gctgtggccc agcgcatagg
     1801 accctatgag gtgctagagc cctccagcga ggaggtggag aagaacctgc gtccgttctc
     1861 caccctggat ctgatgactc ctatactagg ggttgcccct gagtacacca ggcagctgct
     1921 gctggaagga cctgtgcggg tgaaggaggg acgagaaggg aagatggatg tgtacctatt
     1981 cctcttctcc gatgtgctcc tggtaaccaa acctcaacgc aaggcggaca gagccaaggt
     2041 catccgtcct cccctcatgc tggagaaact cgtgtgccga ccactgcgag accccaacag
     2101 cttcctgttg atccacctca cagaattcca gtgtgtctcc agtgccctca ctgtgcactg
     2161 tcccagttct acagagcgtg cacggtggct ggagaagacc cagcatgctc agaccactct
     2221 gcagaagctg aagggtgagc agtacattca acagaagagg gagctgctgg ccctctatcg
     2281 caaccagggc acggagtccc ccagcaccag gccctccaca ccttcgcctt ccccggagga
     2341 ctcccagagc agtgccgagg ggaggacttt agagttcgcc atcatccccc gcctggtggt
     2401 gactgaggat gcagatgaag acacgccctc catgccggat gacgcctcag actctggcta
     2461 cagtaccttg attccaagct ccccgaagga ttctcattcc cccctgaaca ggcttcggtc
     2521 caaggctctt cgtagagacc ctcgcctcac cttctccacc ctggaactca gagacgtccc
     2581 cttgcgcccc cagcctcccg acccccaagc tccccagcgg cgaagcgccc cagagttgcc
     2641 agaagggatc ctaagaggag gcagtctacc caggagagcc ccaccaatct ggtcggagga
     2701 ggaagacgag actttggcat cagggaatgt ggtggtagaa accctacaca gggcgcaacg
     2761 tcggagtccg ctcccccact ctcccactca tactgactct gctggggaaa gcccctggga
     2821 gtcctcagat gaagatgagg ggcttctatc ccctgaactc cgcccccgct ccctacggga
     2881 ggacatgctc agggagatca gggaggagct ggccaaccaa aggatcgatg gagcctcgga
     2941 accggaatca gggaatggca agccccggag gctgactcta gcccagctgc agagaatgag
     3001 ggtcccccac atcatacagc tggacacgcc cctgtctaca tcagaagtat aaaagggtgg
     3061 gcgcgccgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     3121 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     3181 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3241 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     3301 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     3361 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3421 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     3481 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3541 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3601 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3661 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3721 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3781 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3841 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3901 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3961 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     4021 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     4081 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     4141 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     4201 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     4261 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4321 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4381 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4441 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4501 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4561 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4621 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4681 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4741 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4801 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4861 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4921 ttgctggcct tttgctcaca tgtt
//
