LOCUS       sequence-223871-        5236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 5' tagging with alpha actinin 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5236)
  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 5236)
  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: pME-actn2
FEATURES             Location/Qualifiers
     source          1..5236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2793..2892)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2909..2928)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2910..2928)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2933..2949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3051..3070)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3062..3871
                     /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(3129..3148)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4760..4777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4936..4963)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5055..5141)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5190..5212
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5204..5221
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5205..5221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct gccaccatga tgaatcagat
      121 cgagctttca gtgccttatg ataacggata tgaaatagac gagtacatga tgcaggagga
      181 tgaatgggac cgggatttac tgctggatcc agcatgggaa aagcaacaaa ggaagacgtt
      241 cactgcctgg tgtaactctc acctgagaaa agctgggacg cagatcgaga acatagagga
      301 ggactttaga aatggcctca aactaatgct gctgcttgaa gtcatctcag gtgagaggtt
      361 acccaagcct gacagaggga agatgcgttt tcacaagata gctaatgtca ataaagccct
      421 ggatttcatc accagtaaag gtgtgaaact ggtctccatt ggagctgaag aaattgtgga
      481 tggtaatgtg aaaatgactc ttggaatgat ctggacaatc atccttcgct tcgccattca
      541 ggatatctct gttgaagaaa cttcggccaa agaaggactt ctgctgtggt gtcagagaaa
      601 gactgccccg taccgaaatg tcaatgttca aaacttccat gtcagctgga aggatggatt
      661 ggcgctgtgc gctctcatcc acaggcacag gcctgacctc attgactacg ccaagctcaa
      721 caaggatgat ccactgggta atttaaatct ggcatttgac attgctgaaa aacatctgga
      781 catccccaaa atgctggatg cagaggacat cctgagcacc cctaaacctg atgagagggc
      841 tattatgaca tacgtgtcct gcttttacca tgcatttgct ggagctgagc aggctgaaac
      901 tgctgcgaac aggatctgta aggtgcttgg agtgaatcag gaaaatgaga agttgatgga
      961 ggattatgaa aggctggcta gtgagctgtt ggagtggatc agacgtacca ttccttggct
     1021 ggagaatcgt gcagctgaga agaccatggt tgagatgcag aggaagctgg aagattttcg
     1081 agattaccgc aggatgcaca aaccacccaa agtgcaggag aaatgtcagc tggaaatcag
     1141 cttcaacacg ctgcagacca aactccgcat cagcaaccgg cctgcgttta tgccctctga
     1201 gggcaaaatg gtgtcggaca tagcgagtgc gtggcaaggg ctggaacaag cagagaaggg
     1261 ctatgaggaa tggctgctga ctgagatcag gaggctggaa agagtggagc atctagctga
     1321 gaaattcaga cagaaagcta ctaaccatga gtcctgggct acaggtaagg aagagatgtt
     1381 gacccggaag gactacgagt cagcgtctct aatggaagtg cgagctttgc tgcggaaaca
     1441 cgaagcgttt gagagtgacc tttctgcaca ccaggacaga gttgagcaga tcgctgctat
     1501 tgctcaggag ctcaatgagc tggactatca tgatgttgca tccgtgaatc aaaggtgtca
     1561 aagtatttgt gacctctggg accatctggg cacgctcacc cagaaacgca gagaggcttt
     1621 ggagaggaca gagaaacttt tggagacggt ggatcagttg tttctggaat atgcaaagag
     1681 atctgcaccc tttaataact ggatggaagg agctatggaa gacttgcagg acatgttcat
     1741 tgtgcacaca attgaggaag tccagactct gatagctgct catgagcagt ttaaggccac
     1801 ccttcctgag gcagatgcag agagacaggc gatccttggc attcagcagg aagtcctgaa
     1861 gatcttccag agttatggca tccgtggaga tcttacaaat ccctacagca ctattactgc
     1921 agaagaaatt gccatcaaat gggacaaggt gaagaagctg gtgccccagc gtgacagtgc
     1981 actgcaggag gagttagcga gacagcatgc taacgaaaga ctcagacgcc agtttgctgc
     2041 ccaagccaac ctcatcggac cctggataca gactaggatg gaggaaattg gtcattgttc
     2101 tatggcgatg ggcggaactc tggaggatca gatgacccaa ctgaagcagt atgagcatgt
     2161 gatcgtcagc tacaaatcca acatagaccg acttgaggga gatcaccaac tcattcagga
     2221 gtcacttatc tttgacaaca aacacactaa ctacactatg gagcatgctc gcgttggctg
     2281 ggagctccta ttgaccacga ttgctcgcac aattaatgaa atcgagactc aaattttgac
     2341 ccgtgatgca aaagggatca gccaggagca gatgcatgag ttccggtcct ctttcaacca
     2401 ttttgatcgg aagaagaacg ggggcatgga ttctgatgac tttagggcct gtctgatctc
     2461 gatgggttat gatctgggag aggtggagtt tgctcggatc atgatgctgg tggatccaaa
     2521 tgcttcaggc aatgtgtcgt tccagtcctt cattgatttc atgactagag agacagcaga
     2581 cacagacaca gctgaacagg tcattgcttc cttcaagatc ctcgctgcag acaagccgta
     2641 tattctgtca gatgagttga ggcgagagct tcctccagac caggccgagt actgtatcag
     2701 tcgaatgcct ccttactgtg gccccggggc cctgcctgga gccttggact acaccgcctt
     2761 ctccaccgcc ctatatggag aaagtgacct tgacccagct ttcttgtaca aagttggcat
     2821 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2881 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2941 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     3001 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     3061 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     3121 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3181 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3241 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     3301 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3361 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3421 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3481 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3541 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3601 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3661 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3721 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3781 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3841 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3901 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3961 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4681 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4741 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     4801 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     4861 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     4921 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     4981 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
     5041 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     5101 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
     5161 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
     5221 tcttaagctc gggccc
//
