LOCUS       sequence-222015-        4690 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Multisite gateway vector for 3' tagging with junctophilin 1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4690)
  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 4690)
  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-jph1a
FEATURES             Location/Qualifiers
     source          1..4690
                     /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(2176..2271)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2308..2327)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2309..2327)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2332..2348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2332..2348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2450..2469)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2461..3270
                     /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(2528..2547)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3417..4005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3906..3925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4159..4176
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4335..4362)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4454..4540)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4589..4611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4603..4620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4604..4620
                     /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 gtggtgacgg gcggacggtt cgacttcgac gatggaggca cttactgtgg cggatgggag
      181 gacgggaaag cccacggaca cggcatctgc acgggaccca agggtcaggg cgagtactcc
      241 gggtcctggt cccacgggtt tgaggtagtc ggggtctaca cctggcccag cgggaacacg
      301 taccagggct actggtcgca ggggaagcgc cacgggctgg gtgttgagac caaggggaaa
      361 tggctttatc gcggggagtg gagtcacggg ttcaagggtc gctatggagt tcggcagagc
      421 accaacacac ctgctagata tgatgggacg tggagtaatg gactgcagga tggatatgga
      481 gtcgaaacct atggggatag tggcacttat caaggccagt ggatgggggg catgaggcat
      541 ggctatggcg tccgtcagag cgtgccttat ggaatggcca ccatcatccg ttctccgctc
      601 cgcacctccc tggcctctct aagaagtgtg caaagcaacg gcgccgttct ccaggacaca
      661 gtaccagaca ctccagttgg aagccgtggt ggtttcgtgc tgaattttca cagcgacacc
      721 gaagttgtca ccggtaagaa gaagggcctg ttccgtcgcg ggtccctctt cgggagcctc
      781 cgccagctgc gtaaatctga ctccaaaacg tccatctcta gcaagcgaag ctcagcacgt
      841 agcgatgcca ccatgagccg catcagctcc agcgacgcca actccaccat cagctatgga
      901 gaaggagagg tgcctgatga ttacacgcca atggaggacc acgtggatgc caccaccact
      961 gagtcctaca tgggcgaatg gaagaacgat aagcgcaatg ggtttggggt cagtgagaga
     1021 tccaacgggc tgaagtatga aggggaatgg atgaacaaca agaggcatgg atacgggtgc
     1081 accatattcc ctgatggaac caaagaggag gggaagtata aaaataacgt gttggtgcgc
     1141 gggataagga agcagctcat tcctcttaaa aaccataaaa cgaaggaaaa agtggaccgg
     1201 gctgtggagg gggctcgaag gtcggcagct atcgccagga cgaaagtgga aatagcagtt
     1261 tcaaggacgt cccacgcacg gactaagagt gaagcagcag accaggccgc tctcgcagca
     1321 tgtcaggagt cagatttagc ccgagctgtg gcttgtgaat tatctccaag cttccatcaa
     1381 ccaggactgg aatacatcaa gcagaagttc agtgagccgg tggaggagat taaagaagaa
     1441 gcacccaaag aagaggagaa gaagtcttca ggtagccctt tctataggcg cgggacgact
     1501 ccgagccact ctccagccca aagcccgacc ccatcccctc cccgcacctc tcagaaaagc
     1561 caagcgcaga accccagcat tagccgcaag atcagcaaag acaacagcgt aacggcccgc
     1621 aagatcagca gagacagcgc ttctgtagcc cgcaagatca gcaaagaaga aagagcctcc
     1681 attgtggccg aaatcaccaa agtatccgta cctcctcctc ctcttcctcc tcctcctcct
     1741 cctgcagttc agactatcaa gctgcaggat gtcccacctc caaagcctcc taaagcactt
     1801 gaacccgtgg ccaaacctgc gggcacaggt aacgggcagc ttcatactgc ctatcacagt
     1861 tattatgtga agccggtgaa gcagttgcca ccgccagagg agcccgaaga tgaggagcct
     1921 gaaatcacac agtccagtct ggccaggatg cctccacctc cgaaaacgct caacacaccc
     1981 actcccaaac cagaggtcaa gatcaggaaa caagagtcta ttaaaccaaa gagcttagct
     2041 gaaaccaaga aggccagctt ggacctggtt gaagaagcaa cggtggaaga atctggtcct
     2101 aactcaattc tggttgccat ggtgatgctt ttgaatattg gattggcaat catttttgtc
     2161 catttcctga catgacaact ttattataca aagttggcat tataaaaaag cattgcttat
     2221 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tggagctcca
     2281 tggtagcgtt aacgcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct
     2341 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2401 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2461 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2521 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2581 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2641 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2701 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2761 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2821 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2881 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2941 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     3001 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     3061 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     3121 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     3181 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3241 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3301 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3361 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3481 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3541 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3601 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3661 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3721 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3781 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3841 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3901 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3961 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4021 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4081 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4141 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4201 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4261 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     4321 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     4381 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     4441 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     4501 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     4561 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     4621 cttaagctcg ggccctgcag ctctagagct cgaattctac aggtcactaa taccatctaa
     4681 gtagttggtt
//
