LOCUS       sequence-222042-        3943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD1b-delta-EH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3943)
  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 3943)
  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-dEH
FEATURES             Location/Qualifiers
     source          1..3943
                     /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(1429..1524)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1561..1580)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1562..1580)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1585..1601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1585..1601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1703..1722)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1714..2523
                     /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(1781..1800)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2670..3258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3159..3178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3412..3429
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3588..3615)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3707..3793)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3842..3864
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3856..3873
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3857..3873
                     /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 ttgattaaca actttattat
     1441 acaaagttgg cattataaaa aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1501 tcagtcaaaa taaaatcatt atttggagct ccatggtagc gttaacgcgg ccgcgatatc
     1561 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     1621 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1681 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     1741 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1801 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     1861 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     1921 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1981 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2041 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2101 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2161 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2221 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2281 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2341 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2401 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2461 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2521 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     2581 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     2641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2821 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2881 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2941 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3001 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3061 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3121 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3181 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3241 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3301 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3361 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3421 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3481 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3541 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     3601 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     3661 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     3721 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     3781 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     3841 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccctg cagctctaga
     3901 gctcgaattc tacaggtcac taataccatc taagtagttg gtt
//
