LOCUS       sequence-222041-        4237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4237)
  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 4237)
  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-EHD1b
FEATURES             Location/Qualifiers
     source          1..4237
                     /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(1723..1818)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1855..1874)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1856..1874)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1879..1895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1879..1895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1997..2016)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2008..2817
                     /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(2075..2094)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2964..3552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3453..3472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3706..3723
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3882..3909)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4001..4087)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4136..4158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4150..4167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4151..4167
                     /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 gtggctttca gttggtctaa caaagatgga aaaaagaaag acccggagtt atttcaaacg
      181 gtttcggacg ggctaaagag actgtacagg accaaacttt tccccttaga ggacgcatac
      241 cgctttcacg acttccactc tccggctctg gaagatgctg atttcgataa caagcccatg
      301 gtacttttgg tgggccaata ctctacaggg aaaaccactt ttattcggca tctcatggag
      361 caggatttcc ctggtatgcg catagggccc gaaccgacca ccgattcatt catagcggtt
      421 atgcacgggg aacaagaagg tctcatcccc ggcaatgctt tagtggttga ccccaagaaa
      481 cccttcagaa aactcaacgc ctttggaaac gcctttctga acaggttcat atgtgctcag
      541 atgccaaacc cagtgctgga cagcatcagc atcatagaca ctcctggaat tctgtccgga
      601 gagaaacaac ggatcagcag aggttatgat tttgctgcgg tgcttgaatg gtttgccgaa
      661 cgggtggacc gaatcattct tctctttgac gcacacaagt tggacatctc ggatgagttc
      721 tccgaggtga tccgagctct gaaaaaccat gaggacaaaa tgcgggtggt gcttaataaa
      781 gctgatcaga tcgggaccca gcagcttatg agggtgtatg gcgccctcat gtggtcactt
      841 gggaaaattg tcaacactcc agaggtcata cgtgtgtata ttggctcttt ctgggctcag
      901 ccgctcctgg ttgcagataa caggaaattg tttgaggcag aggaacaaga ccttttccgt
      961 gacatccagg gacttccacg taatgcagca ctgcgcaaac tgaatgacct tatcaaacgg
     1021 gcccgtctag ccaaagttca agcatacatc atcagctctt tgaagaaaga aatgccaaat
     1081 gtgtttggaa aagagaacaa gaaaaaggag ctcattgcta acctgggtga aatctatgca
     1141 aagattgaaa aagaacatca gatctcccct ggtgattttc ctaaactcag taaaatgcag
     1201 gaagtcctcg ctggacagga cttcactaag tttcaaggat tgaagcccaa gcttttggaa
     1261 tcagtggaag acatgctggc caacgacatt gccaaactca tgaccctcgt ccggcaggag
     1321 gaggcctcca tgccaagcca ggctgtccaa ggtggagcct ttgaaggcac catgaatggc
     1381 cccttcgggc acggctacgg agaaggtgcc ggtgaaggca ttgatgagct ggagtgggta
     1441 gtggccaaag acaaaccgac atacgacgag atcttctaca ctttgtcacc cgtcaatggg
     1501 aaggtgtctg gtgccatggc caagaaggag atggttaagt ccaagctgcc caataatgtt
     1561 ctaggaaaga tctggaagct cgctgacgtg gataaggatg gatacctgga tgacgaggag
     1621 ttcgccctgg ccaaccacct catcaaggtt aaactggaag gtcacgagtt gccagctgat
     1681 cttccagatc accttgtgcc accttcaaaa cgagaacagt aacaacttta ttatacaaag
     1741 ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     1801 aaaataaaat cattatttgg agctccatgg tagcgttaac gcggccgcga tatcccctat
     1861 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1921 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     1981 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2041 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2101 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2161 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2221 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2281 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     2341 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2401 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2461 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2521 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2581 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2641 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2701 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2761 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     2821 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     2881 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2941 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3001 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3061 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3121 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3181 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3241 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3301 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3361 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3421 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3481 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3541 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3601 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3661 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3721 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3781 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3841 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3901 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3961 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     4021 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     4081 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     4141 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cctgcagctc tagagctcga
     4201 attctacagg tcactaatac catctaagta gttggtt
//
