LOCUS       sequence-224022-        3296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab4b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3296)
  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 3296)
  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-rab4b
FEATURES             Location/Qualifiers
     source          1..3296
                     /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(682..777)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(814..833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(815..833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(838..854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(838..854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(956..975)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             967..1776
                     /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(1034..1053)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1923..2511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2412..2431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2665..2682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2841..2868)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2960..3046)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3095..3117
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3109..3126
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3110..3126
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggtgtcag agacgtacga tttcctgttc aagttcctgg
       61 tgattggcag cgccgggact gggaaatcat gcctccttca ccagttcata gagaacaagt
      121 tcaaacagga ctccaatcac accatcggcg ttgagttcgg ctccagggtt gtcaatgttg
      181 gtggcaaaac agtcaaactg cagatctggg acacggctgg acaggagcgc tttaggtcag
      241 tgacccgcag ttactatcgt ggagcagcag gagctcttct cgtgtatgac atcacaagtc
      301 gggagaccta caatgctctg actaactggt tgacagacgc aaggacactg gccagtccca
      361 acatcgtcat catcctgtgc ggaaataaga aggatctggg cgcagaccgt gaagtcactt
      421 tcctagaggc atctcgcttc gcccaggaaa atgagctgat gtttttagag acgagtgctt
      481 tgactgggga gaatgtcgag gaagcctttc ttaaatgcgc tcgcagcatc ctcaacaaga
      541 ttgaatcagg tgagctggac ccagagcgga tgggttctgg gattcagtat ggcgacgcgt
      601 ccctgcggca gatccgacag ccccggggct ctgctgcaca gaccaagcag cagtgtaact
      661 gttagggctc tgaagaccct tcaactttat tatacaaagt tggcattata aaaaagcatt
      721 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga
      781 gctccatggt agcgttaacg cggccgcgat atcccctata gtgagtcgta ttacatggtc
      841 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
      901 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
      961 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     1021 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1081 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1141 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     1201 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     1261 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     1321 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1381 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     1441 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     1501 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     1561 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     1621 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     1681 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     1741 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     1801 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     1861 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2041 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2281 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2581 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     2641 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     2701 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     2761 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
     2821 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
     2881 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     2941 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     3001 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     3061 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
     3121 gccagtctta agctcgggcc ctgcagctct agagctcgaa ttctacaggt cactaatacc
     3181 atctaagtag ttggttcata gtgactgcat atgttgtgtt ttacagtatt atgtagtctg
     3241 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctc
//
