LOCUS       sequence-206477-        2884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with the CaaX domain from 
            Kras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2884)
  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 2884)
  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: p3E-CaaX(tK)
FEATURES             Location/Qualifiers
     source          1..2884
                     /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    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    807..941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(831..850)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     885..904
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(993..1088)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1125..1144)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1126..1144)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1149..1165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1149..1165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1267..1286)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1278..2087
                     /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(1345..1364)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2234..2822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2723..2742
                     /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtgggaa agaagaagaa gaagaagagc aagaccaagt
      781 gcgtgatcat gtgatcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
      841 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      901 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
      961 attttatgtt tcaggttcag ggggaggtgt ggcaacttta ttatacaaag ttggcattat
     1021 aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     1081 cattatttgg agctccatgg tagcgttaac gcggccgcga tatcccctat agtgagtcgt
     1141 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     1201 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     1261 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     1321 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1381 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1441 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     1501 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1561 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     1621 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1681 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1741 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1801 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     1861 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     1921 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     1981 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     2041 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     2101 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     2161 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     2221 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2281 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2341 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2401 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2461 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2521 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2581 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca
     2641 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2701 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2761 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2821 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2881 tgtt
//
