LOCUS       sequence-222037-        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"
     protein_bind    complement(1..125)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4329..4424)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4461..4480)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4462..4480)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4485..4501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4485..4501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4603..4622)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4614..5423
                     /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(4681..4700)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5570..6158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6059..6078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6312..6329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6488..6515)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6607..6693)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6742..6764
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6756..6773
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6757..6773
                     /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 gtggtcgact taagacgaat attacagatg actcctggga aaggaggctc tcagagctcg
      181 gagtctgagc agccgtctgc tgatatgaat aatgagtcct cagagggctt tatttgtccc
      241 cagtgtatga aatcccataa ctctgctgag gagctcttca aacattatga ggtctttcat
      301 gaaccaggag accagccgtc cccctttagc cccggcaggg aggatttaac attacttcga
      361 caggaaatcc aagacttaca ggcttcactg aaggaggaac gctggttttc tgaggaattg
      421 aaaaaagaat tagacaaagt acaaggacga atagcaacta ataaaggctt acagagcgat
      481 ggtacaagtg gagaagatgc agagctcgag atgagactca atgaatcgga gacagagaag
      541 ttcaacatca aacagatgaa ggacctgttt gaacaaaaag ctgctcagct tgcgaccgag
      601 atagttgaca tcaaatctcg ctacgatgag gagaaaagta tgagagaagc tttggagcag
      661 agactggcca acttaaacca agatatgcaa aaagaaaaac aggagaagga gaaactgtcg
      721 gcagagctgc tccagaggcc gggtgtggaa gatgtggagg tgttgaagaa agagctggtg
      781 cgggtgcaga cgctcatgga cagcatgacc cgagagcgag aggaggagtc agagcgcctc
      841 aagggccaat atgagcagct gcaggccaac ttcactacct cagagaaaac gattgctcaa
      901 ttaaaggctg aactggagaa gggtcctcaa gaagcagcgg tttacacaca acagatccac
      961 caattacaga gcagcttgaa caacctccag cagcaatccc aggctttgtc agagaaatta
     1021 tctcgtaaag agaaggagaa tcaggaactg gaggagcgtc tgggtcacga acaggcctct
     1081 aagaagagtc ttcaggctaa tctgcatcag aaagagctgg agctccagga gagtcgagca
     1141 cgggtgtctt caggagaagc agctctcagt cgagcacagg ctgaattaac tgaacgtggg
     1201 gaagaggctg cacgactcag acgagagctg ggcgagctgg agaagaacca gcaggagctg
     1261 aaggcagagc ggaaacaact tcaacagcag agggaggata aagaaaacca gggcctgcag
     1321 caacagagtg agatcagcca gctgcatgcg aagctgctgg aagcggagcg tcaggtgggg
     1381 gagctgcagg gtcgtctgaa agagcagaga cagctgtctg gagagaagct caaagacaga
     1441 gaacagcagg cagcagatct ccagctcaaa ctctcccgct tagaggaaga gttaaaggag
     1501 agctccacca agtccactga cctgcagcat cagctggaca aatccaaaca gcagcatcaa
     1561 gaactacaaa cacttcaaca gagcactaat ggcaaactta gagaagcaca gaatgacctg
     1621 gagcaggtgt tgcgtcagat cggcgacaaa gaccagaaga tccagaatct tgaagctctg
     1681 ctccagaaga gtaaggaaag tgtgagccag ctggagaccg agagagagga tctgtgtgcc
     1741 aagattcagg ctggagaagg agaagctgct ttgctgaacc aattacaaga gaaaaatcac
     1801 tcgctacaag aacagatcac gcagcttaca gacaagctta agaaccagtc ggagagccac
     1861 aagcaggctc aggacaacct tcatgaacag gtgcaggagc agaagacaca cctgcgttca
     1921 gctcaagacc gctgtcaggg tctggagacc accgtcactg aactcaacac acagctcaca
     1981 gagagcagag agaagatcgc tcagctagac actcagttga aagccaagac tgagatgctg
     2041 ttgtctgctg aagcagcaaa aaatgcccag agagctgact tggagagcca cctagagact
     2101 gctcagaatg cattgcagga taagcagcaa gagctcagta aagtgcaggc tcagctggag
     2161 gagcaggccc gcaggctgac ggagaagcag gagcagtgct ctcagctgga gaacggcctg
     2221 aaggacagca gagacaaact gatgactgca gaacagagga tagagacgct ccagacccag
     2281 accaagaaag ccgaggttga gttgactgag ctccgctccg gaagagaaca ggctcagaaa
     2341 gagcagtctt ctcttaagaa gcagatcagt gagctggaga tgaagaccaa agagttgaac
     2401 cgacttttag attccgagaa acaaggagct tccacccagc aagaggagct aaagaagaaa
     2461 agctcagctc tcaccgaaac aagacaaaag ctggagaaag cagagcagga gcgtggagcc
     2521 ctgcaggcca acctggataa actgtctcag gagggtcaga agcagcaggc agagttggac
     2581 aagaaggtgc agggtctgag ttcagatctc cagaaaactc agggggagaa agaagcgctg
     2641 gtaaaagaag tggcgacagt gaaagaggct ctgagtaaag cttctaaagc cctgaaggag
     2701 agccagactc aattagacaa ggagaagaag ggcagcaaga ccactttgga ggagaaggaa
     2761 aaggtttttc aaaaagtaca acaagagctt cagaagaact cagaggccac ttctaaagag
     2821 ctcagcaatg tcaaaggtct gctggagaaa tcgatagagg cagaaaagag tcttcaatct
     2881 cagctgactg cactgaacgc tcagctgaag cagtgtcagg atacactgaa ggagaaagaa
     2941 aagagtgaac agcagctgca ggcagagctg aagaccaggc aaggctcctt cagtcaggaa
     3001 gtgaagaaac tgaagacaca ggtgtctgag ctgcaggctt cccttgctaa aaagactgaa
     3061 gaggaagcca agcaaaagga gcaaatcaca gcactgtcta aagatctgag ctcagagaag
     3121 aatcgctgtg cagagctgca gaaaaccagt gacggcacca aagagagtct caccgccctt
     3181 cagtctgatt actatggcaa agagtctgag ctctctgctg tacgccagga ccttaaggtg
     3241 tgcgaggaga agttggttct tgctcaggag gagctggtga ccaacaggaa ccagctcagt
     3301 gcttacgaaa cccaaatcca ggagctgaag acgggccaca cagcgatgga gatagacctg
     3361 agcaagagag acgagaagat caaacagcaa gtggagactc tgcagaagct ccagaaacaa
     3421 cagggacaaa cagaggagca gctgaagaag gagaaagctc aatgcgagga gctgagagag
     3481 agtcaaagcg ccctggaaaa ggacaaaaac aaactgtctg ctgatctcaa aacactggct
     3541 gagaagaatg agaaggagct gaaggacttg caggcagcaa agcagttgtt gatccagcaa
     3601 aaggtggaga tgcaggggaa ggtagaggcg gctcaagcat ctgtggagaa agaacagaga
     3661 gagcatcaga agaccagaga ctccatcaaa cagaatgagc agcagctcaa ggctgagacc
     3721 aacaaaataa agcagcagct ggcatcagaa gtgaaggcta aagaggagat ggtgcggcag
     3781 agggaagagg ctgaggtgaa gatgtccatg caggtgacgg cactgaatga gaacgtggcc
     3841 acgctgaagc gagagtggca gagcagccag aggcgggtgg gtgagctgga gaaacagacg
     3901 gatgagctga gaggagagat tgccgtgctg gaagcaacag tccagaacaa ccaagacgaa
     3961 agacgtgcac tacttgagag gtgcgtccag ggtgaagggg agatggagaa gcttcaggct
     4021 aagatgttgg agatgcgcag gaagttggac gacactactg ccgccatgca ggagcttggt
     4081 cgagagaacc agtccttaca gataaaacag tcacaatccc tcacacggaa gtggacagaa
     4141 gaccatgagg tgcagaactg catggcttgt gggaaaggct tctccgtcac tgtcagaaag
     4201 caccactgca gacactgcgg gaacatcttc tgtgcagagt gctctgcccg caacgccctc
     4261 acaccctcct ccaagaagcc cgttcgagtg tgtgacaact gctttgatga gctccagggc
     4321 tgactagtca actttattat acaaagttgg cattataaaa aagcattgct tatcaatttg
     4381 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggagct ccatggtagc
     4441 gttaacgcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     4501 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4561 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     4621 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     4681 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     4741 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     4801 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     4861 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     4921 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     4981 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     5041 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     5101 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     5161 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     5221 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     5281 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     5341 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     5401 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     5461 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     5521 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5701 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6181 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6241 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6301 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     6361 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6421 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     6481 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     6541 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     6601 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     6661 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt
     6721 taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc
     6781 tcgggccctg cagctctaga gctcgaattc tacaggtcac taataccatc taagtagttg
     6841 gtt
//
