LOCUS       sequence-223867-        2602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 5' tagging with the actin binding motif
            Lifeact.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2602)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 2602)
  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: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: pME-Lifeact
FEATURES             Location/Qualifiers
     source          1..2602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      101..110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             107..157
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     protein_bind    complement(159..258)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(275..294)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(276..294)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(299..315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(299..315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(417..436)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             428..1237
                     /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(495..514)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1384..1972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1873..1892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2126..2143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2302..2329)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2421..2507)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2556..2578
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2570..2587
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2571..2587
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct gccaccatgg gcgtggccga
      121 cctgatcaag aagttcgaga gcatcagcaa ggaggagaac ccagctttct tgtacaaagt
      181 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
      241 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
      301 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
      361 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
      421 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
      481 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
      541 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
      601 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
      661 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
      721 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
      781 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
      841 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
      901 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
      961 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1021 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     1081 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     1141 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     1201 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     1261 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     1321 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1381 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1441 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1501 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1561 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1621 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1681 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1741 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1801 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1861 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1921 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1981 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2041 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2101 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2161 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2221 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     2281 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
     2341 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
     2401 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     2461 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
     2521 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
     2581 ggccagtctt aagctcgggc cc
//
