LOCUS       sequence-222122-        3181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative ARF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3181)
  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 3181)
  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-ARF1-DN
FEATURES             Location/Qualifiers
     source          1..3181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..25)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(567..662)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(699..718)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(700..718)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(723..739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(723..739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(841..860)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             852..1661
                     /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(919..938)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1808..2396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2297..2316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2550..2567
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2726..2753)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2845..2931)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2980..3002
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2994..3011
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2995..3011
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggtgggaa acatattcgc aaacctcttt aaagtcttcg
       61 gcaagaagga gatgagaatt cttatggtgg gactcgacgc agcagggaag aacacgattc
      121 tgtataaatt aaagttgggg gaaattgtga ccaccatccc aacaatcggt tttaatgttg
      181 aaacagtgga gtacaagaat attagtttta cagtttggga cgtcggcggt caagataaga
      241 ttcgaccatt gtggcgtcat tatttccaga acacacaagg tctgatcttt gtggttgaca
      301 gtaatgaccg tgagcgtgtg aatgaggcgc gagaggagct gatgaggatg ttggctgagg
      361 atgagctcag agaagcagtt ctgctggtgt ttgccaacaa acaggacctg ccgaacgcca
      421 tgaacgccgc cgaaatcacg gacaagctgg gtctgcactc actccggcac agaaactggt
      481 acattcaggc cacgtgtgcc acaagcggcg acggcctcta tgaaggactc gactggttgt
      541 ccaatcagct gaaaaaccag aaatgacaac tttattatac aaagttggca ttataaaaaa
      601 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
      661 ttggagctcc atggtagcgt taacgcggcc gcgatatccc ctatagtgag tcgtattaca
      721 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
      781 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
      841 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
      901 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
      961 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1021 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1081 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1141 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1201 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1261 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1321 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     1381 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     1441 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     1501 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     1561 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     1621 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     1681 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     1741 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1801 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1861 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1921 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1981 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2041 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2101 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2161 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2221 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2281 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2341 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2401 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2461 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2521 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2581 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2641 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     2701 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     2761 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     2821 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     2881 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     2941 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa
     3001 cgacggccag tcttaagctc gggccctgca gctctagagc tcgaattcta caggtcacta
     3061 ataccatcta agtagttggt tcatagtgac tgcatatgtt gtgttttaca gtattatgta
     3121 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     3181 c
//
