LOCUS       sequence-206524-        4111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with Pacsin2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4111)
  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 4111)
  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-Pacsin2
FEATURES             Location/Qualifiers
     source          1..4111
                     /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..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             1230..1241
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(1810..1827)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2220..2315)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2352..2371)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2353..2371)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2376..2392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2376..2392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2494..2513)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2505..3314
                     /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(2572..2591)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3461..4049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3950..3969
                     /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 aaagtggctt cggatttcaa cgactctctg attgacgtgt
      781 ccagcgacag cttctgggag gtgggaaact acaagcgttc agtgaagcga gtggacgatg
      841 ggaaccgtct gtgtaacgac ctgatgagct gcctgcacga gcgcgcacgc atcgagaaga
      901 gctacagcac acagctcacc gactgggcca agcgctggag acagaccatc gagaaaggcc
      961 ctcagtacgg gacggtggag cgggcgtggt gcgctctgat gaccgaggcc gagaaggtga
     1021 gcgatctgca catggaggtg aaggccgctc tgatggcgga ggacttcgag aagatcaaga
     1081 actggcagaa ggacgcctac cacaagcaga taatcggggg cttcaaggag accaaagagg
     1141 cggaggacgg ctttcgcaag gctcagaaac cctgggccaa gaagctgaag gagatggaga
     1201 ccatgaagaa ggcgtaccac acggcctgca aagaggagaa aacagccacc agcagagaga
     1261 actccagcaa actggacaac agcaaccccg aagcccagaa gaagctgcag gagaaggtgg
     1321 agaagtgcca gcaggaggtg cagaagacga aggagcgcta cgagaagtct ctggaagatc
     1381 tggacaaggt gacgccgcag tacatggaga acatggagca ggtgttcgag cagtggcagc
     1441 agttcgaggg aaagagactg agcttcttca aggaggtcct gctggaggtc aagcagcacc
     1501 tcgacctctc ctccaatcac aagtacacca ccgtctacca cacgctacag gacactatcc
     1561 agggagcaga tccacaagag gacctcaagt ggttccgctc aaaccacgga cccggcatga
     1621 ccatgaactg gccccagttt gaggaatggt ccatggacct gaaccggacc ctcagccgcc
     1681 gggaaacgaa gcgcagacct gctgagggag tgaccctgac agcgatcagt cacaatcctg
     1741 agcagagctc cagagtcagc agtgtgcctg cggtcagtag ttcagaagcg gcatgcttaa
     1801 acccgtttga tgatgatgat gatgatgtag atgaggatga agagccggcg gtcacacaca
     1861 cccacagcag cccggtcaaa catgaggagg ctgtagcaca aactgcagca gtagagaagg
     1921 ctcccgctcc agctgccatc actgctgctg atgatgatga tgatgaagag tcaggaaacc
     1981 ccttctcctc ctcgtcagcc aatgggaatc cgtttgagga ggagccgtcc acggagatgt
     2041 gtgtgccggt gcgcgctgtc tatgactatg agggacagga gcaggacgag ctcagcttca
     2101 aagcaggaga tgagctggtg aagctgagtg aggaggacga gcagggctgg tgtaaggggc
     2161 gtctgagcga cggcactgtt ggtctgtatc ctgccaacta tgtggaggat ctgcagtgac
     2221 aactttatta tacaaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga
     2281 acaggtcact atcagtcaaa ataaaatcat tatttggagc tccatggtag cgttaacgcg
     2341 gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     2401 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     2461 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     2521 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     2581 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     2641 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     2701 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     2761 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     2821 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     2881 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2941 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     3001 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     3061 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3121 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3181 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3241 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     3301 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     3361 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     3421 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3481 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3541 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3601 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3661 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3721 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3781 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3841 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3901 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3961 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4021 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4081 tggccttttg ctggcctttt gctcacatgt t
//
