LOCUS       sequence-206497-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 5' tagging with early endosome 
            associated 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  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 6843)
  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-EEA1
FEATURES             Location/Qualifiers
     source          1..6843
                     /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..748)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4952..5047)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5084..5103)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5085..5103)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5108..5124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5108..5124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5226..5245)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5237..6046
                     /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(5304..5323)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6193..6781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6682..6701
                     /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 aaagtggtcg acttaagacg aatattacag atgactcctg
      781 ggaaaggagg ctctcagagc tcggagtctg agcagccgtc tgctgatatg aataatgagt
      841 cctcagaggg ctttatttgt ccccagtgta tgaaatccca taactctgct gaggagctct
      901 tcaaacatta tgaggtcttt catgaaccag gagaccagcc gtcccccttt agccccggca
      961 gggaggattt aacattactt cgacaggaaa tccaagactt acaggcttca ctgaaggagg
     1021 aacgctggtt ttctgaggaa ttgaaaaaag aattagacaa agtacaagga cgaatagcaa
     1081 ctaataaagg cttacagagc gatggtacaa gtggagaaga tgcagagctc gagatgagac
     1141 tcaatgaatc ggagacagag aagttcaaca tcaaacagat gaaggacctg tttgaacaaa
     1201 aagctgctca gcttgcgacc gagatagttg acatcaaatc tcgctacgat gaggagaaaa
     1261 gtatgagaga agctttggag cagagactgg ccaacttaaa ccaagatatg caaaaagaaa
     1321 aacaggagaa ggagaaactg tcggcagagc tgctccagag gccgggtgtg gaagatgtgg
     1381 aggtgttgaa gaaagagctg gtgcgggtgc agacgctcat ggacagcatg acccgagagc
     1441 gagaggagga gtcagagcgc ctcaagggcc aatatgagca gctgcaggcc aacttcacta
     1501 cctcagagaa aacgattgct caattaaagg ctgaactgga gaagggtcct caagaagcag
     1561 cggtttacac acaacagatc caccaattac agagcagctt gaacaacctc cagcagcaat
     1621 cccaggcttt gtcagagaaa ttatctcgta aagagaagga gaatcaggaa ctggaggagc
     1681 gtctgggtca cgaacaggcc tctaagaaga gtcttcaggc taatctgcat cagaaagagc
     1741 tggagctcca ggagagtcga gcacgggtgt cttcaggaga agcagctctc agtcgagcac
     1801 aggctgaatt aactgaacgt ggggaagagg ctgcacgact cagacgagag ctgggcgagc
     1861 tggagaagaa ccagcaggag ctgaaggcag agcggaaaca acttcaacag cagagggagg
     1921 ataaagaaaa ccagggcctg cagcaacaga gtgagatcag ccagctgcat gcgaagctgc
     1981 tggaagcgga gcgtcaggtg ggggagctgc agggtcgtct gaaagagcag agacagctgt
     2041 ctggagagaa gctcaaagac agagaacagc aggcagcaga tctccagctc aaactctccc
     2101 gcttagagga agagttaaag gagagctcca ccaagtccac tgacctgcag catcagctgg
     2161 acaaatccaa acagcagcat caagaactac aaacacttca acagagcact aatggcaaac
     2221 ttagagaagc acagaatgac ctggagcagg tgttgcgtca gatcggcgac aaagaccaga
     2281 agatccagaa tcttgaagct ctgctccaga agagtaagga aagtgtgagc cagctggaga
     2341 ccgagagaga ggatctgtgt gccaagattc aggctggaga aggagaagct gctttgctga
     2401 accaattaca agagaaaaat cactcgctac aagaacagat cacgcagctt acagacaagc
     2461 ttaagaacca gtcggagagc cacaagcagg ctcaggacaa ccttcatgaa caggtgcagg
     2521 agcagaagac acacctgcgt tcagctcaag accgctgtca gggtctggag accaccgtca
     2581 ctgaactcaa cacacagctc acagagagca gagagaagat cgctcagcta gacactcagt
     2641 tgaaagccaa gactgagatg ctgttgtctg ctgaagcagc aaaaaatgcc cagagagctg
     2701 acttggagag ccacctagag actgctcaga atgcattgca ggataagcag caagagctca
     2761 gtaaagtgca ggctcagctg gaggagcagg cccgcaggct gacggagaag caggagcagt
     2821 gctctcagct ggagaacggc ctgaaggaca gcagagacaa actgatgact gcagaacaga
     2881 ggatagagac gctccagacc cagaccaaga aagccgaggt tgagttgact gagctccgct
     2941 ccggaagaga acaggctcag aaagagcagt cttctcttaa gaagcagatc agtgagctgg
     3001 agatgaagac caaagagttg aaccgacttt tagattccga gaaacaagga gcttccaccc
     3061 agcaagagga gctaaagaag aaaagctcag ctctcaccga aacaagacaa aagctggaga
     3121 aagcagagca ggagcgtgga gccctgcagg ccaacctgga taaactgtct caggagggtc
     3181 agaagcagca ggcagagttg gacaagaagg tgcagggtct gagttcagat ctccagaaaa
     3241 ctcaggggga gaaagaagcg ctggtaaaag aagtggcgac agtgaaagag gctctgagta
     3301 aagcttctaa agccctgaag gagagccaga ctcaattaga caaggagaag aagggcagca
     3361 agaccacttt ggaggagaag gaaaaggttt ttcaaaaagt acaacaagag cttcagaaga
     3421 actcagaggc cacttctaaa gagctcagca atgtcaaagg tctgctggag aaatcgatag
     3481 aggcagaaaa gagtcttcaa tctcagctga ctgcactgaa cgctcagctg aagcagtgtc
     3541 aggatacact gaaggagaaa gaaaagagtg aacagcagct gcaggcagag ctgaagacca
     3601 ggcaaggctc cttcagtcag gaagtgaaga aactgaagac acaggtgtct gagctgcagg
     3661 cttcccttgc taaaaagact gaagaggaag ccaagcaaaa ggagcaaatc acagcactgt
     3721 ctaaagatct gagctcagag aagaatcgct gtgcagagct gcagaaaacc agtgacggca
     3781 ccaaagagag tctcaccgcc cttcagtctg attactatgg caaagagtct gagctctctg
     3841 ctgtacgcca ggaccttaag gtgtgcgagg agaagttggt tcttgctcag gaggagctgg
     3901 tgaccaacag gaaccagctc agtgcttacg aaacccaaat ccaggagctg aagacgggcc
     3961 acacagcgat ggagatagac ctgagcaaga gagacgagaa gatcaaacag caagtggaga
     4021 ctctgcagaa gctccagaaa caacagggac aaacagagga gcagctgaag aaggagaaag
     4081 ctcaatgcga ggagctgaga gagagtcaaa gcgccctgga aaaggacaaa aacaaactgt
     4141 ctgctgatct caaaacactg gctgagaaga atgagaagga gctgaaggac ttgcaggcag
     4201 caaagcagtt gttgatccag caaaaggtgg agatgcaggg gaaggtagag gcggctcaag
     4261 catctgtgga gaaagaacag agagagcatc agaagaccag agactccatc aaacagaatg
     4321 agcagcagct caaggctgag accaacaaaa taaagcagca gctggcatca gaagtgaagg
     4381 ctaaagagga gatggtgcgg cagagggaag aggctgaggt gaagatgtcc atgcaggtga
     4441 cggcactgaa tgagaacgtg gccacgctga agcgagagtg gcagagcagc cagaggcggg
     4501 tgggtgagct ggagaaacag acggatgagc tgagaggaga gattgccgtg ctggaagcaa
     4561 cagtccagaa caaccaagac gaaagacgtg cactacttga gaggtgcgtc cagggtgaag
     4621 gggagatgga gaagcttcag gctaagatgt tggagatgcg caggaagttg gacgacacta
     4681 ctgccgccat gcaggagctt ggtcgagaga accagtcctt acagataaaa cagtcacaat
     4741 ccctcacacg gaagtggaca gaagaccatg aggtgcagaa ctgcatggct tgtgggaaag
     4801 gcttctccgt cactgtcaga aagcaccact gcagacactg cgggaacatc ttctgtgcag
     4861 agtgctctgc ccgcaacgcc ctcacaccct cctccaagaa gcccgttcga gtgtgtgaca
     4921 actgctttga tgagctccag ggctgactag tcaactttat tatacaaagt tggcattata
     4981 aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     5041 attatttgga gctccatggt agcgttaacg cggccgcgat atcccctata gtgagtcgta
     5101 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     5161 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     5221 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     5281 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     5341 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     5401 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     5461 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     5521 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     5581 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     5641 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     5701 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     5761 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     5821 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     5881 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     5941 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     6001 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     6061 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     6121 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     6181 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6241 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6301 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     6361 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6421 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6481 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6541 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     6601 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6661 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6721 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6781 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6841 gtt
//
