LOCUS       sequence-206650-        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"
     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"
     protein_bind    complement(6618..6713)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6750..6769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6751..6769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6774..6790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6774..6790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6892..6911)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6903..7712
                     /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(6970..6989)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7859..8447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8348..8367
                     /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 aaagtgggtc gactggaaga aggaggagaa accaactcaa
      781 ttcctgaaca aagtggcatc agtgtcattc caagcccagt tgttcagaga agaaaaatag
      841 gctcaactcg aagacctcta aaagggaata atgtgactgc agagtcagat gtgcttcaaa
      901 aagccacttt agctgatgca aaattagaaa aagacaatgt atctgaaaat actgagagcc
      961 tttcagctgt caccgattct aatcttttac ttcaacatca tggcactgaa ggagcaggag
     1021 aaacagactc aattcctaaa caaagtggca tcagcatcat ccctggagat cttcaaagaa
     1081 aggagacaaa cttagagtca agtccagtta ttcagagaag aaaaatgggc tcaactcgaa
     1141 gacctctcaa aggaaataaa ggacaaagaa aaggacgaga gcttcatgat gaaaatgaaa
     1201 catttaacag tgaatttttg cttgatgtac agaaagaaac attcggtttg gaggatgaac
     1261 cctgtccaga tgaatctaca ttacaaaaca taaaagatgg agctgaagaa caggagaagg
     1321 aactgtgtgt gaagaatgat gctagtcttg ttggagagtc tgaatctgct ctcattcagt
     1381 catcatacgg ttcagattta gcgtctggag aaaccactgc attgagtgtc tgtaacatca
     1441 cagaaccaca aaccctcaca aaaactcacg aagaggcctg ggaaatagtt gggggtgttg
     1501 aaagagcaac agtagaagaa gacacagact tgcaatcaaa aaccgagtca caaaaccaaa
     1561 catgtgcctc aattagtgaa gcagaagaaa acatccatga tgtaaatgtt ttagtcagag
     1621 aagctgaagt tactggagaa tcagttatat tggatagtaa tgtgactgca gagtcagatg
     1681 tgattcaaaa agacacttta gctgatgcag aattagaaac attgcaagtt tctgaaaata
     1741 cagaggatgt ttctgctgtg actgatactg attttctagt gcaacatcat ggcactgaac
     1801 gagcagaagg gacagacaca atttctgtac aaagtggcat caacatcatt cctagagagt
     1861 ctcacggaga agaggcgaac tcaaagtcga gtccagttgt tcagaggaga aaaatgggct
     1921 caactcgaag acctctcaaa ggaaataaag gacaaagaaa aggacaagag cttgatgatg
     1981 aaaaggaaac atttcactgt gaatttttgc ttgatgtaga aaaagaaaca tttagtttgg
     2041 agaatgaacc ctgtccagat gaatctacat tacaaaacat gaaagatgga gctgaagaac
     2101 aggagaagga aatatgtctg gagaatgatg ttggtcttgt tgaagagtct gaatctgctc
     2161 tcattcagtc atatagttca gatttagtgt ctgaagaatc cactgcattg agtgtctgta
     2221 acatcacaga atcacaaacc ctcacaaaaa ttcaagaaga gtcctgggaa atagttggag
     2281 atgttgaaag agaatcagta gatggagata cagacttaca attagaaaca gagtcacaaa
     2341 accaaacatg tgactcaacc agtaaagcag aacaaaacat ccatgatgta aatgtattag
     2401 tcagagaagc tgaaggtatt ggggaatcag ttaaagtgga cagcaatgtg attgcagagt
     2461 cagatgtgct tcaaaaagcc actttagctg atccaaaatt ggaaacattg cacgtatctg
     2521 aaaatactaa ggatgtttct gctgtcacgg attctgatct tctagtgcaa catgaaccaa
     2581 ctgtagaagc aggagaaaca gacacggttc ctgaaaaaag tggaatcagc atcattcctg
     2641 gagattctca caacgaagag gcaaactcaa agtcaagtcc agttgttcag aggagaaaaa
     2701 tgggctcaac tcgaagacct ctcaaaggaa ataaaggaca aagaaaagga cgagagcttc
     2761 aagatgaaaa tgagacactt aataatgaaa ttaggtttga tgtagagaaa gaaacatttg
     2821 gtttggagaa tgaaccctgt ccagatgaat ctgcattaca aaacataaaa gttggggctg
     2881 aagaacagga gaaggaaata tgtgtgaaga atgatgctgg tgttgttgaa gagtatgaat
     2941 ctgatctcat tcagtcattt tatggttcgg atttagtgtc tgaagaatcc actgcattaa
     3001 gtgtctgcca catcgaagaa gcacaaactc tcacaattag tcatgaacaa tttcagaata
     3061 atcctggaga tgcagaaaga gcagcagtag ttgaagatgc tgccgatact gaagatactt
     3121 cagaaactga agctacagtt acaatgaaga gcactacaaa tactgcatca catgtttttc
     3181 aaacgtctcc tagttcaagc acagatgcat ttaatatatc tacaaatgtc gaggaagata
     3241 cttcaggaga cacagacaca attttcaagc agagtaacat caacgttgtt catgaagata
     3301 tttatagagg ggaagctcac ccaaagttaa gtccagttgt tcagaagaga aaaatgggct
     3361 caactcgaag acctctcagg gggaacaaag gacaaaaaaa agataatgat gaaaaggaaa
     3421 tctcatttct ggaaaatgaa cccaatccag atgagcttat atacactgca atgggtatca
     3481 gtgatatttc agaagctcaa acccacgtaa aagttcacga agagtcccag gacaaccatg
     3541 aagttgttga aaaagcaaca gtaggggaaa gcacacaatc acaaacagaa aacaaagtga
     3601 gtgtgttagt cagagaagct gaagttcctg aaaatacagt tgtgattaat acaactgagg
     3661 agtcgtattt gtttcaatac gtctctgatt caaaactgac catatctgac tatgaacatg
     3721 aggctccagt aattccaaac ataatttcca aacaaagtga cttcagcatt cctggagatt
     3781 ctcacaaaga agaggaaaac tcacaattaa gtccagttgt tcaaaagaga aaaatgggct
     3841 caactcgaag acctctcaaa gggaataagg ggcaaggaaa gcagagagaa gaatccaagg
     3901 taaagaatag agataatgag gcagggtcag aagaggaaaa cacagaaaca cattcaaaag
     3961 accaggaaca aaatgtggca aaagacaata tgaatataat agataatgtc aatgaagaaa
     4021 aaaaggcaac aaaggcattg gatacatcca tgcaggaact atgtgaacaa gagcaaagat
     4081 cagatgtatc acaaacacat gctgatacat cacaggatga aaccagctca catttattgg
     4141 acatcccaga aagaactgta gattttgtta gggagtctac gagtcagcct gatcttgttc
     4201 tgtctgatgt gttgcttatg acagaagctc aaacgtattt gtcatcagaa gataatgtca
     4261 tgggtaacag ttcaatatct ataacagagg tacatgaaat gcatgcagac tccaatacat
     4321 ccaagagaaa aatggggtca actagaaaga gaggaaagag gatggtgaat gaagaggaag
     4381 aggaagctga aaatacaatt ggttctggag ccacaagttc caaggagaca ttaactccta
     4441 aaaaattaat tgcagaatct tctgtacata aaccactgtt tagttcatat gttattaacg
     4501 aagaccatga cagtaaatca gatgtgcaga ctgagctctc agatcaagat tatctaaatt
     4561 ctaccataaa caccgaacca atgaaagaaa gagaaagttc tccagatgtt gttgctgaaa
     4621 tggtttctga ctttgctttg ggacatgttg acactgaagt agttcctgag caaagggtca
     4681 ttcctgaaga gccacataaa gatcaagcaa acctgaatcc agttgttcaa aaacgaaaaa
     4741 tggggtcaac acgtaaaact cttagaggga acaaagggca gagaaaagag gaagaatcga
     4801 aagaaattaa tgaaggaatg acacaggatc ggagtagaga actacatttg gcaagtgaaa
     4861 gtaaacttgg tgaagaggtt caacaaaacc aatctcttat taatatgaca aacaataaag
     4921 aagctaagaa accggaggat gaattggatg aaatgccttc agcatcaagc atctctcaat
     4981 tcacagaagc atcctcagaa actctcagtt cctcatatct taaatcagaa aagttagatt
     5041 caaaagatga gaatgattat catgatactt gtgaacatga aaatgcccat cagatttgcg
     5101 ctgatgctac caaaagtcta cagtcggaat ctgttcccat tgagaggaga aggaagatgg
     5161 gctcgacccg gaaaaatatt ggaagagaag gaagaatgca aagaaagcga gatggatatg
     5221 aggacacaga aatcatagac acaaatctgc agatgtctca cgaccctgaa atgagctcat
     5281 caggtataac tgcagaagaa caaacaactg aagattactc caatcagtca gaggattcac
     5341 ttcttggtga aatacagcga atgctaaaaa caacttctca aagtgcatcc aattttgaat
     5401 tgattattga tgagccgaaa tcaaattcac cagaaggctc agaaatcaga tctctttcac
     5461 catcacagag cttgacaact ctaaatgatc caaactctcc agaaagacga cggcgaaaaa
     5521 tgggttcaac tcgcaagaac cctaaacggc agttcaaagc agaaagagaa gatgaagaca
     5581 aagtagatca gaagacagag tcaaacaaag aagaaataac ggaggggtcg ctagtgcata
     5641 atatcacaga agaccctcca aataaagagt atcttgatgt tcccagtgca cacacaagca
     5701 gccagcaaga agagagcgat agccctgttg ttcaagaaag tgcgtcacca tcaactaaaa
     5761 ggaagtttgg gtccagacga gctaataagg gcaaacttgg taggcttgag gcatctgatg
     5821 gtatgactga atcagaagat ggagatcata aaccagatgg ctctgctgtt gaggacagac
     5881 agcacaggga tggttcaatg ggttgtggta cgagcgtcag cttacaacca gaaaatctgc
     5941 cattttccag aaattatgaa gttgctccaa aaggcactgg tgaagagatt caagtcagaa
     6001 aacaggacag aatgacggtc aatgtaaaga acagggattc taaagtggtt cagtttaatg
     6061 tggtcatggt gggaaacagc tgtgttggaa agacgtcctt cataagacgt ttccatgaag
     6121 gccagttcac cgaggattac cgctccacta ttggtgttga cactttcgta cagactgttg
     6181 agctgcccga cagaactgtt aaactgcaga tatgggacac agcaggtcaa gagaggtttc
     6241 acagcatcac cacgcaagtt ttccacaagg ctgaggggct tctgctcatg tatgagatca
     6301 catgttcaaa gagtttcatt tctatacgag attggatctc tcaagcccgg gaaagggctc
     6361 aagatgatgt cgtcatgatg ttgctcggaa acaagaatga ttcagttaat cgtgaagtcc
     6421 agattcagga aggagcagac ctggccagag agtataacat acatttcatg gagtgcagtg
     6481 ctgcaaacgg ggctaatgtg tcagaatcta tgaggactct tgctgagttg ttagtgcagc
     6541 gcaagagaaa aagggaggaa cacacaacgc tgagaagaga accacaacag aagaaatctg
     6601 gatgctgtta accgcggcaa ctttattata caaagttggc attataaaaa agcattgctt
     6661 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggagctc
     6721 catggtagcg ttaacgcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     6781 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     6841 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     6901 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     6961 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     7021 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     7081 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     7141 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     7201 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     7261 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     7321 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     7381 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     7441 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     7501 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     7561 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     7621 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     7681 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     7741 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     7801 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7861 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7921 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7981 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     8041 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8101 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8161 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8221 acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga
     8281 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8341 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8401 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8461 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
