LOCUS       sequence-222119-        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"
     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"
     RBS             607..618
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(1187..1204)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1597..1692)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1729..1748)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1730..1748)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1753..1769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1753..1769)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1871..1890)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1882..2691
                     /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(1949..1968)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2838..3426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3327..3346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3580..3597
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3756..3783)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3875..3961)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4010..4032
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4024..4041
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4025..4041
                     /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 gtggcttcgg atttcaacga ctctctgatt gacgtgtcca gcgacagctt ctgggaggtg
      181 ggaaactaca agcgttcagt gaagcgagtg gacgatggga accgtctgtg taacgacctg
      241 atgagctgcc tgcacgagcg cgcacgcatc gagaagagct acagcacaca gctcaccgac
      301 tgggccaagc gctggagaca gaccatcgag aaaggccctc agtacgggac ggtggagcgg
      361 gcgtggtgcg ctctgatgac cgaggccgag aaggtgagcg atctgcacat ggaggtgaag
      421 gccgctctga tggcggagga cttcgagaag atcaagaact ggcagaagga cgcctaccac
      481 aagcagataa tcgggggctt caaggagacc aaagaggcgg aggacggctt tcgcaaggct
      541 cagaaaccct gggccaagaa gctgaaggag atggagacca tgaagaaggc gtaccacacg
      601 gcctgcaaag aggagaaaac agccaccagc agagagaact ccagcaaact ggacaacagc
      661 aaccccgaag cccagaagaa gctgcaggag aaggtggaga agtgccagca ggaggtgcag
      721 aagacgaagg agcgctacga gaagtctctg gaagatctgg acaaggtgac gccgcagtac
      781 atggagaaca tggagcaggt gttcgagcag tggcagcagt tcgagggaaa gagactgagc
      841 ttcttcaagg aggtcctgct ggaggtcaag cagcacctcg acctctcctc caatcacaag
      901 tacaccaccg tctaccacac gctacaggac actatccagg gagcagatcc acaagaggac
      961 ctcaagtggt tccgctcaaa ccacggaccc ggcatgacca tgaactggcc ccagtttgag
     1021 gaatggtcca tggacctgaa ccggaccctc agccgccggg aaacgaagcg cagacctgct
     1081 gagggagtga ccctgacagc gatcagtcac aatcctgagc agagctccag agtcagcagt
     1141 gtgcctgcgg tcagtagttc agaagcggca tgcttaaacc cgtttgatga tgatgatgat
     1201 gatgtagatg aggatgaaga gccggcggtc acacacaccc acagcagccc ggtcaaacat
     1261 gaggaggctg tagcacaaac tgcagcagta gagaaggctc ccgctccagc tgccatcact
     1321 gctgctgatg atgatgatga tgaagagtca ggaaacccct tctcctcctc gtcagccaat
     1381 gggaatccgt ttgaggagga gccgtccacg gagatgtgtg tgccggtgcg cgctgtctat
     1441 gactatgagg gacaggagca ggacgagctc agcttcaaag caggagatga gctggtgaag
     1501 ctgagtgagg aggacgagca gggctggtgt aaggggcgtc tgagcgacgg cactgttggt
     1561 ctgtatcctg ccaactatgt ggaggatctg cagtgacaac tttattatac aaagttggca
     1621 ttataaaaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     1681 aaatcattat ttggagctcc atggtagcgt taacgcggcc gcgatatccc ctatagtgag
     1741 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1801 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1861 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1921 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1981 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2041 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2101 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     2161 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     2221 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2281 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2341 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2401 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     2461 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2521 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2581 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2641 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2701 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2761 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2821 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2881 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2941 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3001 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3061 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3121 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3181 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3241 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3301 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3361 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3421 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3481 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3541 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3601 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3661 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3721 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3781 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3841 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3901 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3961 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     4021 cgttgtaaaa cgacggccag tcttaagctc gggccctgca gctctagagc tcgaattcta
     4081 caggtcacta ataccatcta agtagttggt t
//
