LOCUS       sequence-223992-        8509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab44.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8509)
  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 8509)
  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-rab44
FEATURES             Location/Qualifiers
     source          1..8509
                     /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"
     protein_bind    complement(5995..6090)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6127..6146)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6128..6146)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6151..6167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6151..6167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6269..6288)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6280..7089
                     /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(6347..6366)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7236..7824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7725..7744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7978..7995
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(8154..8181)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(8273..8359)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     8408..8430
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8422..8439
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8423..8439
                     /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 gtgggtcgac tggaagaagg aggagaaacc aactcaattc ctgaacaaag tggcatcagt
      181 gtcattccaa gcccagttgt tcagagaaga aaaataggct caactcgaag acctctaaaa
      241 gggaataatg tgactgcaga gtcagatgtg cttcaaaaag ccactttagc tgatgcaaaa
      301 ttagaaaaag acaatgtatc tgaaaatact gagagccttt cagctgtcac cgattctaat
      361 cttttacttc aacatcatgg cactgaagga gcaggagaaa cagactcaat tcctaaacaa
      421 agtggcatca gcatcatccc tggagatctt caaagaaagg agacaaactt agagtcaagt
      481 ccagttattc agagaagaaa aatgggctca actcgaagac ctctcaaagg aaataaagga
      541 caaagaaaag gacgagagct tcatgatgaa aatgaaacat ttaacagtga atttttgctt
      601 gatgtacaga aagaaacatt cggtttggag gatgaaccct gtccagatga atctacatta
      661 caaaacataa aagatggagc tgaagaacag gagaaggaac tgtgtgtgaa gaatgatgct
      721 agtcttgttg gagagtctga atctgctctc attcagtcat catacggttc agatttagcg
      781 tctggagaaa ccactgcatt gagtgtctgt aacatcacag aaccacaaac cctcacaaaa
      841 actcacgaag aggcctggga aatagttggg ggtgttgaaa gagcaacagt agaagaagac
      901 acagacttgc aatcaaaaac cgagtcacaa aaccaaacat gtgcctcaat tagtgaagca
      961 gaagaaaaca tccatgatgt aaatgtttta gtcagagaag ctgaagttac tggagaatca
     1021 gttatattgg atagtaatgt gactgcagag tcagatgtga ttcaaaaaga cactttagct
     1081 gatgcagaat tagaaacatt gcaagtttct gaaaatacag aggatgtttc tgctgtgact
     1141 gatactgatt ttctagtgca acatcatggc actgaacgag cagaagggac agacacaatt
     1201 tctgtacaaa gtggcatcaa catcattcct agagagtctc acggagaaga ggcgaactca
     1261 aagtcgagtc cagttgttca gaggagaaaa atgggctcaa ctcgaagacc tctcaaagga
     1321 aataaaggac aaagaaaagg acaagagctt gatgatgaaa aggaaacatt tcactgtgaa
     1381 tttttgcttg atgtagaaaa agaaacattt agtttggaga atgaaccctg tccagatgaa
     1441 tctacattac aaaacatgaa agatggagct gaagaacagg agaaggaaat atgtctggag
     1501 aatgatgttg gtcttgttga agagtctgaa tctgctctca ttcagtcata tagttcagat
     1561 ttagtgtctg aagaatccac tgcattgagt gtctgtaaca tcacagaatc acaaaccctc
     1621 acaaaaattc aagaagagtc ctgggaaata gttggagatg ttgaaagaga atcagtagat
     1681 ggagatacag acttacaatt agaaacagag tcacaaaacc aaacatgtga ctcaaccagt
     1741 aaagcagaac aaaacatcca tgatgtaaat gtattagtca gagaagctga aggtattggg
     1801 gaatcagtta aagtggacag caatgtgatt gcagagtcag atgtgcttca aaaagccact
     1861 ttagctgatc caaaattgga aacattgcac gtatctgaaa atactaagga tgtttctgct
     1921 gtcacggatt ctgatcttct agtgcaacat gaaccaactg tagaagcagg agaaacagac
     1981 acggttcctg aaaaaagtgg aatcagcatc attcctggag attctcacaa cgaagaggca
     2041 aactcaaagt caagtccagt tgttcagagg agaaaaatgg gctcaactcg aagacctctc
     2101 aaaggaaata aaggacaaag aaaaggacga gagcttcaag atgaaaatga gacacttaat
     2161 aatgaaatta ggtttgatgt agagaaagaa acatttggtt tggagaatga accctgtcca
     2221 gatgaatctg cattacaaaa cataaaagtt ggggctgaag aacaggagaa ggaaatatgt
     2281 gtgaagaatg atgctggtgt tgttgaagag tatgaatctg atctcattca gtcattttat
     2341 ggttcggatt tagtgtctga agaatccact gcattaagtg tctgccacat cgaagaagca
     2401 caaactctca caattagtca tgaacaattt cagaataatc ctggagatgc agaaagagca
     2461 gcagtagttg aagatgctgc cgatactgaa gatacttcag aaactgaagc tacagttaca
     2521 atgaagagca ctacaaatac tgcatcacat gtttttcaaa cgtctcctag ttcaagcaca
     2581 gatgcattta atatatctac aaatgtcgag gaagatactt caggagacac agacacaatt
     2641 ttcaagcaga gtaacatcaa cgttgttcat gaagatattt atagagggga agctcaccca
     2701 aagttaagtc cagttgttca gaagagaaaa atgggctcaa ctcgaagacc tctcaggggg
     2761 aacaaaggac aaaaaaaaga taatgatgaa aaggaaatct catttctgga aaatgaaccc
     2821 aatccagatg agcttatata cactgcaatg ggtatcagtg atatttcaga agctcaaacc
     2881 cacgtaaaag ttcacgaaga gtcccaggac aaccatgaag ttgttgaaaa agcaacagta
     2941 ggggaaagca cacaatcaca aacagaaaac aaagtgagtg tgttagtcag agaagctgaa
     3001 gttcctgaaa atacagttgt gattaataca actgaggagt cgtatttgtt tcaatacgtc
     3061 tctgattcaa aactgaccat atctgactat gaacatgagg ctccagtaat tccaaacata
     3121 atttccaaac aaagtgactt cagcattcct ggagattctc acaaagaaga ggaaaactca
     3181 caattaagtc cagttgttca aaagagaaaa atgggctcaa ctcgaagacc tctcaaaggg
     3241 aataaggggc aaggaaagca gagagaagaa tccaaggtaa agaatagaga taatgaggca
     3301 gggtcagaag aggaaaacac agaaacacat tcaaaagacc aggaacaaaa tgtggcaaaa
     3361 gacaatatga atataataga taatgtcaat gaagaaaaaa aggcaacaaa ggcattggat
     3421 acatccatgc aggaactatg tgaacaagag caaagatcag atgtatcaca aacacatgct
     3481 gatacatcac aggatgaaac cagctcacat ttattggaca tcccagaaag aactgtagat
     3541 tttgttaggg agtctacgag tcagcctgat cttgttctgt ctgatgtgtt gcttatgaca
     3601 gaagctcaaa cgtatttgtc atcagaagat aatgtcatgg gtaacagttc aatatctata
     3661 acagaggtac atgaaatgca tgcagactcc aatacatcca agagaaaaat ggggtcaact
     3721 agaaagagag gaaagaggat ggtgaatgaa gaggaagagg aagctgaaaa tacaattggt
     3781 tctggagcca caagttccaa ggagacatta actcctaaaa aattaattgc agaatcttct
     3841 gtacataaac cactgtttag ttcatatgtt attaacgaag accatgacag taaatcagat
     3901 gtgcagactg agctctcaga tcaagattat ctaaattcta ccataaacac cgaaccaatg
     3961 aaagaaagag aaagttctcc agatgttgtt gctgaaatgg tttctgactt tgctttggga
     4021 catgttgaca ctgaagtagt tcctgagcaa agggtcattc ctgaagagcc acataaagat
     4081 caagcaaacc tgaatccagt tgttcaaaaa cgaaaaatgg ggtcaacacg taaaactctt
     4141 agagggaaca aagggcagag aaaagaggaa gaatcgaaag aaattaatga aggaatgaca
     4201 caggatcgga gtagagaact acatttggca agtgaaagta aacttggtga agaggttcaa
     4261 caaaaccaat ctcttattaa tatgacaaac aataaagaag ctaagaaacc ggaggatgaa
     4321 ttggatgaaa tgccttcagc atcaagcatc tctcaattca cagaagcatc ctcagaaact
     4381 ctcagttcct catatcttaa atcagaaaag ttagattcaa aagatgagaa tgattatcat
     4441 gatacttgtg aacatgaaaa tgcccatcag atttgcgctg atgctaccaa aagtctacag
     4501 tcggaatctg ttcccattga gaggagaagg aagatgggct cgacccggaa aaatattgga
     4561 agagaaggaa gaatgcaaag aaagcgagat ggatatgagg acacagaaat catagacaca
     4621 aatctgcaga tgtctcacga ccctgaaatg agctcatcag gtataactgc agaagaacaa
     4681 acaactgaag attactccaa tcagtcagag gattcacttc ttggtgaaat acagcgaatg
     4741 ctaaaaacaa cttctcaaag tgcatccaat tttgaattga ttattgatga gccgaaatca
     4801 aattcaccag aaggctcaga aatcagatct ctttcaccat cacagagctt gacaactcta
     4861 aatgatccaa actctccaga aagacgacgg cgaaaaatgg gttcaactcg caagaaccct
     4921 aaacggcagt tcaaagcaga aagagaagat gaagacaaag tagatcagaa gacagagtca
     4981 aacaaagaag aaataacgga ggggtcgcta gtgcataata tcacagaaga ccctccaaat
     5041 aaagagtatc ttgatgttcc cagtgcacac acaagcagcc agcaagaaga gagcgatagc
     5101 cctgttgttc aagaaagtgc gtcaccatca actaaaagga agtttgggtc cagacgagct
     5161 aataagggca aacttggtag gcttgaggca tctgatggta tgactgaatc agaagatgga
     5221 gatcataaac cagatggctc tgctgttgag gacagacagc acagggatgg ttcaatgggt
     5281 tgtggtacga gcgtcagctt acaaccagaa aatctgccat tttccagaaa ttatgaagtt
     5341 gctccaaaag gcactggtga agagattcaa gtcagaaaac aggacagaat gacggtcaat
     5401 gtaaagaaca gggattctaa agtggttcag tttaatgtgg tcatggtggg aaacagctgt
     5461 gttggaaaga cgtccttcat aagacgtttc catgaaggcc agttcaccga ggattaccgc
     5521 tccactattg gtgttgacac tttcgtacag actgttgagc tgcccgacag aactgttaaa
     5581 ctgcagatat gggacacagc aggtcaagag aggtttcaca gcatcaccac gcaagttttc
     5641 cacaaggctg aggggcttct gctcatgtat gagatcacat gttcaaagag tttcatttct
     5701 atacgagatt ggatctctca agcccgggaa agggctcaag atgatgtcgt catgatgttg
     5761 ctcggaaaca agaatgattc agttaatcgt gaagtccaga ttcaggaagg agcagacctg
     5821 gccagagagt ataacataca tttcatggag tgcagtgctg caaacggggc taatgtgtca
     5881 gaatctatga ggactcttgc tgagttgtta gtgcagcgca agagaaaaag ggaggaacac
     5941 acaacgctga gaagagaacc acaacagaag aaatctggat gctgttaacc gcggcaactt
     6001 tattatacaa agttggcatt ataaaaaagc attgcttatc aatttgttgc aacgaacagg
     6061 tcactatcag tcaaaataaa atcattattt ggagctccat ggtagcgtta acgcggccgc
     6121 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     6181 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     6241 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     6301 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     6361 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     6421 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     6481 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     6541 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     6601 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     6661 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     6721 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     6781 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     6841 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     6901 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     6961 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     7021 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     7081 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     7141 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     7201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7381 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7981 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8041 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8101 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     8161 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     8221 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     8281 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     8341 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     8401 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccctgcagc
     8461 tctagagctc gaattctaca ggtcactaat accatctaag tagttggtt
//
