LOCUS       sequence-222012-        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"
     protein_bind    1..96
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    148..282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(185..204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     239..258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    342..465
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(536..553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(536..552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(545..567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      616..702
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      794..821
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(980..997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1151..1739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1231..1250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1886..2695)
                     /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     2609..2628
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2687..2706
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2808..2824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2808..2824
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2829..2848
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2829..2847
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat
       61 gcttttttat aatgccaact ttgtataata aagttgccac acctccccct gaacctgaaa
      121 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
      181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
      241 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctacgtaat acgatcacat
      301 gatcacgcac ttggtcttgc tcttcttctt cttcttcttt cccactttgt acaagaaagt
      361 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa
      421 acagactaca taatactgta aaacacaaca tatgcagtca ctatgaacca actacttaga
      481 tggtattagt gacctgtaga attcgagctc tagagctgca gggcccgagc ttaagactgg
      541 ccgtcgtttt acaacgtcgt gactgggaaa acatccatgc tagcgttaac gcgagagtag
      601 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt
      661 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
      721 aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
      781 catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
      841 cctggctagc ggtacgcgta ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      961 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1021 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1081 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1561 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg taactcacgt
     1801 taagggattt tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg ctctgccagt
     1861 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     1921 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1981 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     2041 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     2101 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     2161 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     2221 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     2281 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     2341 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt tttccgggga
     2401 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     2461 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     2521 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaagcgat
     2581 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2641 catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg ctcataacac
     2701 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     2761 cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgccag gaaacagcta
     2821 tgaccatgta atacgactca ctatagggga tatcgcggcc gcgttaacgc taccatggag
     2881 ctcc
//
