LOCUS       sequence-222038-        3733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 5' tagging with Bin1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3733)
  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 3733)
  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-Bin1b
FEATURES             Location/Qualifiers
     source          1..3733
                     /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(1219..1314)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1351..1370)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1352..1370)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1375..1391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1375..1391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1493..1512)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1504..2313
                     /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(1571..1590)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2460..3048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2949..2968
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3202..3219
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3378..3405)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3497..3583)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3632..3654
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3646..3663
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3647..3663
                     /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 gtggtggcag aggttgggaa aggagtcaac gccgggaaac tggccagtaa tgtacagaag
      181 aggttaaccc gagcacaaga gaaggtgatg cagaaactgg gaaaggcgga tgaaaccaaa
      241 gacacagcct ttgaggagca agtagccaac tttaacaaac agatgaatga gggcaccaaa
      301 ctgcagaaag acttgaaagc gtatttgctg gcagtcaaaa caatgcacga gtgttccaag
      361 cgtctgcacg actgtctggc agaaatgtat gaccctgagt ggtttggcaa agaggaggtg
      421 gactccatcg cagaggacac tgacctcctg tggcaagatt tccacgagaa gcttttggac
      481 aatgctgtaa tttccatgga cgcatacttg gctcagtttc cagatataaa ggctcgcatt
      541 gcaaagcgtg aaaggaagct ggtggacttt gacagtgcca gacatcattt tgcctctgta
      601 caaaaaggca agaaaaagga cgaggccaaa attgccaagg ctgaggaaca cttgggcaga
      661 gctcagaaag tgtttgagga gatcaattat gatctccagg aggaacttcc caccctgtgg
      721 aacagtcgta ttggcctcta tgttaacaca ttccggagtg tgtctggcct ggaggaaaca
      781 tttcacaggg acatgggcaa actgaaccag aatctgagtg atattatgac caaactggaa
      841 gagcagcgtg ttgccaaaaa aggtgtcacg acagcaagca ctgggaagag tgacgatgca
      901 gcaaaccaca atcaggagac gtcagatgca cctaaatctc catcaaagaa tgaagcccca
      961 acggtgacaa atggctcctc agatacagag cttcctccat ctgttctctt taaggtgaaa
     1021 acaatgcacg actacagtgc aacagacagt gatgaacttg acctgaaggc tggtgacatt
     1081 gtgcttgtgc ttccctttgc caacccagat gaagcggatg atggctggtt aatgggtgta
     1141 aaagagtccc actggcttca gaacaaaaac ctcctagcaa aaggagtctt ccctgaaaac
     1201 ttcacccaga aactgtgaca actttattat acaaagttgg cattataaaa aagcattgct
     1261 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggagct
     1321 ccatggtagc gttaacgcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata
     1381 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1441 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1501 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1561 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1621 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1681 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1741 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1801 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1861 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1921 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1981 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2041 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2101 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2161 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2221 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2281 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2341 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2401 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2461 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2521 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2581 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2641 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2701 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2761 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2821 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2881 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2941 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3001 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3061 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3121 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3181 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3241 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3301 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3361 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3421 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3481 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3541 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3601 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3661 agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac taataccatc
     3721 taagtagttg gtt
//
