LOCUS       sequence-206506-        4270 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD2b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4270)
  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 4270)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-EHD2b
FEATURES             Location/Qualifiers
     source          1..4270
                     /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(2379..2474)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2511..2530)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2512..2530)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2535..2551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2535..2551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2653..2672)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2664..3473
                     /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(2731..2750)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3620..4208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4109..4128
                     /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 aaagtggtgt cacgctgggg acgtaagaat gtcaagaagg
      781 cccccgaggt gatcaggaca gtgaccgagg gcctgaagtc actctaccga aaaaagctcc
      841 tgcctctgga gcagtattac ggcttccatg acttccactc accgagtctg gaggacgccg
      901 actttgacaa caaacccatg gtgctggtcg ttgggcagta ctccacggga aagaccacct
      961 tcatcaagta tctgcttgaa caggatgtcc ctgggagccg catagggcct gagcccacca
     1021 ccgactgctt caccgctatc atgcacggag acgtagaggg gctcattccc ggaaatgccc
     1081 tcatcgtgga tcccaacaag cccttccgca aactcaaccc cttcggaaac acctttctca
     1141 acagattcca gtgcgctcag ctgcccaacc aggtcctgga gagcatcagc attattgaca
     1201 ccccaggaat cctgtctggg gccaaacaga gagtgagtcg aggatatgat ttcccagcag
     1261 ttctccgctg gtttgctgag cgtgtggacc gaatcatcct tctctttgat gcccacaagt
     1321 tagagatctc agatgaattc tcagaggcta tcggagctct caaaggcaat gaagacaaac
     1381 tccgtgttgt gctgaacaag gctgatatgg ttgataccca gcagcttatg cgggtgtacg
     1441 gagctttaat gtggtccctg gggaaagtgt ttggtactcc tgaggttctg agagtgtaca
     1501 tcggttcctt ctggtctgaa cctctcatgg tgaccgacaa ccgcaagctg ttcgaattgg
     1561 aggaggaaga tctgtttgct gatatccaga atctgccacg taatgcagcc ctacgcaaac
     1621 ttaatgactt ggtcaagagg gcccgtcttg tgagggtaca tgcccatatt atcagttact
     1681 tgaaacaaga gatgccatcg gtcttcagaa aagacaataa aaagaagaac ctgatctacc
     1741 agctcccggt catcttctcc aagatccaac tacaacatca catctctcct ggagacttcc
     1801 ctgactgtgc caagatgcag gagcaactaa tggcacatga cttcaccaag tttaaagccc
     1861 tgaagcccaa ccttatgaat atgctggatg agctgttgtc cagtgacatt gccaaactga
     1921 tgccccttct taggcaggaa gaaatagagg ctggggttca accaggtgtc caaggtggag
     1981 ctttcttggg aacccgtgcc ggaccctttg tggaaggtga cccctttgga gaggttgcga
     2041 atgaaaatgg cgaagtgggt gaagaagatg aggaatgggt ggtgacaaaa gacaagccca
     2101 agtatgatga gatcttctac aacctcgcac cgaatgaagg caaactgagt ggcaccaaag
     2161 ccaaagattg gatggtgagc acaaggctcc ccaactctgt tttgggtcgt atctggaagc
     2221 tttcggatgt ggatcgtgac ggcatgctgg atgatgagga gtttgccctg gccagccatc
     2281 tgattgaagt gaaactggat ggtcatgggc tgccccctga gctccctgcc cgtctcgtgc
     2341 caccatccaa gcgccggcaa aaaggctctg atgcttagca actttattat acaaagttgg
     2401 cattataaaa aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2461 taaaatcatt atttggagct ccatggtagc gttaacgcgg ccgcgatatc ccctatagtg
     2521 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     2581 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     2641 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     2701 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     2761 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     2821 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     2881 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     2941 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     3001 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     3061 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     3121 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     3181 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     3241 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     3301 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     3361 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     3421 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     3481 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     3541 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     3601 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3661 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3721 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3781 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3841 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3901 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3961 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa
     4021 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4081 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4141 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4201 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4261 ctcacatgtt
//
