LOCUS       sequence-222113-        3217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3217)
  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 3217)
  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-Rac1-DN
FEATURES             Location/Qualifiers
     source          1..3217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..124)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(703..798)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(835..854)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(836..854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(859..875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(859..875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(977..996)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             988..1797
                     /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(1055..1074)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1944..2532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2433..2452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2686..2703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2862..2889)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2981..3067)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3116..3138
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3130..3147
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3131..3147
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catagtgact gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
       61 taatttaata tattgatatt tatatcattt tacgtttctc gttcaacttt cttgtacaaa
      121 gtggctcagg ccataaagtg tgtggtcgtc ggggacgggg ctgtgggaaa aaactgcctt
      181 ctgatcagct acaccaccaa tgcttttcct ggggaataca ttcccactgt gtttgataat
      241 tattctgcaa atgtgatggt cgatgggaaa ccagtaaacc tgggattgtg ggatacagca
      301 ggacaagagg attatgacag gcttcgaccg ctttcctacc ctcagacgga tgtgttcctg
      361 atctgcttct ctcttgtgag tccggcctca tttgaaaacg tccgtgcaaa gtggtatccg
      421 gaggtcagac atcattgccc taacactcca ataattcttg ttgggactaa gcttgatttg
      481 cgagatgaca aggacaccat cgaaaagtta aaggagaaga aactcactcc catcacctac
      541 cctcaaggcc ttgctatggc caaagaaata ggagctgtga agtacctgga gtgctctgcc
      601 ttaacccaac gcggccttaa aactgtgttc gatgaagcca tccgtgcagt gctgtgtcca
      661 cctccggtga agaggaggag gagaagatgt ctccttctgt gacaacttta ttatacaaag
      721 ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
      781 aaaataaaat cattatttgg agctccatgg tagcgttaac gcggccgcga tatcccctat
      841 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
      901 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
      961 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     1021 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1081 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     1141 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1201 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1261 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1321 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1381 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1441 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1501 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     1561 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1621 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1681 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1741 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     1801 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     1861 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     1921 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1981 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2041 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2101 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2161 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2221 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2281 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2341 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2401 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2461 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2521 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2581 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2641 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2701 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2761 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2821 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     2881 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     2941 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3001 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3061 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     3121 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cctgcagctc tagagctcga
     3181 attctacagg tcactaatac catctaagta gttggtt
//
