LOCUS       sequence-206505-        4279 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD2a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4279)
  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 4279)
  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-EHD2a
FEATURES             Location/Qualifiers
     source          1..4279
                     /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(2388..2483)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2520..2539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2521..2539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2544..2560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2544..2560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2662..2681)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2673..3482
                     /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(2740..2759)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3629..4217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4118..4137
                     /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 ctgccaaagg aatgaggaac aggggtggta
      781 aatgtctgaa ggcggttcat acagtaaccg aggctttgaa aatcctctat cggcaaaaac
      841 tgctcccact ggagaagtac tatggtttct ccgatttcca ctcatcaagc cttgaagatg
      901 cagactttga caacaaaccc atgatactgg tgttgggcca gtgctctaca ggcaagacca
      961 cttttatcag gtatcttctt gaacaggagt atcctggaag cagagttggc ccagaaccga
     1021 caaccgacag tttcacagca ataatgcatg gagagatgga gggaattatt catggcaatg
     1081 ctttaactgt tgatcccaat aagcctttcc gcaaactcag cccatttgga acaactttcc
     1141 taaatcgatt ccagtgtgta catctgccca atcgggtcct agagagcatc agcatcatcg
     1201 acacaccagg gatcctgtct gtggccaagc gcaaaatgga cagaggttat gactttcctt
     1261 cagttatccg ctggtttgcg gagcacgtag accggatcat cctcctgtgt gatgcacaca
     1321 agctggagat ttctgatgag ttttcccgca ctatactggc attacgaggc aatgaggaca
     1381 agctccgagt ggtgctcaac aaggccgaca tggtggactc gcaggagctg atgcgggtct
     1441 acggcgccct catgtggtct ctcgggaagg tcttctgcac cccggaggtc ctgagggtct
     1501 acatcggctc cttctggacc gctccgatgc gaatcacaga caaccgcaaa ctgtttgagc
     1561 gagaggaggg agatcttttt gcagaccttc agaacctgcc aagaaattct gccctgagaa
     1621 aactcaatga tctggtgaaa cgagcacgcc ttgtaagggc tcatgcttat atcatcagct
     1681 gtctaaaaga cgagatgcct gttgttttct gcagaggaaa gaagcggcga gatctcatct
     1741 atgaccttcc caccatctat tccagaatcc aacagcgcta tcagatttca ccaggagatt
     1801 ttccagactg tgccaagatg caggagcgac tcatgagcca cgacttcaag tttaaaggcc
     1861 tcaagcccaa gctcttagct gggttggatg agctcctaaa caccgacatt gctaagctga
     1921 tgcctcttct acgccaagag gaagaagaag cagctgatca acttttaata caaggaggac
     1981 ctttagggtc tcacaggggc ccgtttattg tggggaaccc atttcatcac tctggatcca
     2041 atggagaaat cgatcaggcg tcaaatgatg aatgggtggt gtcaaaagat aaagcaaagt
     2101 acgacgagat cttttacaac ctctcgcctc atgagggaaa gctgactggg actaaagtga
     2161 aggagtggat gttgcgcact catctgccca gctcggtcct gggacgaatc tggaagctgg
     2221 cagatgtgga ttgtgatggt aagttggacg atgaggagtt tgctttggcc ggtcatctca
     2281 ttgaggtaaa gttggaaggt tttgggcttc ctcatgagct gcccacacac ctgctgccac
     2341 cgtctaaaag acaccgcaag aactctgaga ttaccgacga tgattagcaa ctttattata
     2401 caaagttggc attataaaaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2461 cagtcaaaat aaaatcatta tttggagctc catggtagcg ttaacgcggc cgcgatatcc
     2521 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     2581 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     2641 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     2701 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2761 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     2821 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2881 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     2941 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     3001 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     3061 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     3121 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     3181 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     3241 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     3301 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     3361 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     3421 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     3481 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     3541 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3601 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3661 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3721 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3781 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3841 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3901 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3961 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4021 attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4081 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4141 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4201 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4261 ggccttttgc tcacatgtt
//
