LOCUS       sequence-222013-        8905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with Dysferlin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8905)
  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 8905)
  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-DysF
FEATURES             Location/Qualifiers
     source          1..8905
                     /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"
     CDS             820..831
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(6391..6486)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6523..6542)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6524..6542)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6547..6563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6547..6563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6665..6684)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6676..7485
                     /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(6743..6762)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      7632..8220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8121..8140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8374..8391
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(8550..8577)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(8669..8755)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     8804..8826
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8818..8835
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8819..8835
                     /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 gtggtgttga ggtgtttgct acagagagcg accaacctga gaaatgtaga gaagggggaa
      181 aggcatagcg acccgcttgc tagcatcact ttcagaggga caaaaaagaa gactaaagtc
      241 atcaaaaata atgtcaatcc tgtctggaat gagggctttg agtgggacct gaagggtgtt
      301 cctttagatt cgggagcaga gcttcatgtt gtcgttaaag accatgagaa aatgggcagg
      361 aataggtttc tgggggagtg tcgagttgct cttcgagatg tgctcaattc tccaaacctg
      421 gctgctactt ttaccgtgtc cctggtcgac acaaagagaa acagcactgg ggccacaata
      481 accctgcagg tctcctacat ccctcctcca gggatgacgc ccatctttca gcctcctcca
      541 cagccagagg cccagcacac accggtggag ctggacacag tcacagtttt ctcattggac
      601 actatgggcg aggatgacac tgagagcatg ctgatgatgg aggtggcgga ggagcctgaa
      661 cctgttccag gaccccaggg gcaggacatc ggcgccccta cagcaccacc caaaaaggct
      721 ccacccaact ttaaccaggg actaaagaaa gagaaacgcc aaagcaacaa atcccccctg
      781 tccgacaaag cccaggacct ccaggttcgg ctgcgcatca ttgaaggaag gcagcttcct
      841 ggtgtcaaca tcaaaccagt cgtcaaagtc acagttggag gacagacgaa gagaacccgc
      901 attaaaaagg gcaataatcc attttttgat gagacctttt ttctaaactt ctttgagaaa
      961 ccttcagatc tttttgatga gccaatattc ataaccgtat ttgattccaa gtcactacgg
     1021 aaagactctg tcattggaga attcgagctg gatgttggca tggtttataa tgaacaccga
     1081 catgctattc tcagaaaatg gttgttgctg gcagaccctg atgatctttc tgcaggggcc
     1141 agaggttatt tgaaggtcag catgtttgtg ctggccgcag gagatgaacc tccgatggat
     1201 aagaaagagg gtgtggagga aaaggaggac atagaggcca atctgttgag accagctggt
     1261 ctaaccctga gaggagccac gttcactctg agagtctaca gagcagaaga ccttcctcag
     1321 atgggtgatg ctcttattga cgatgtcaaa catttccttg gctttgacac taatcgaaag
     1381 aacctggtgg atccattggt ggaggtcaac tttgcaggaa aaacggtaag tagtaagatt
     1441 ctggagaaat gcgctaaccc tcagtggaac cagtgcctca ctctgcccat caggtttcca
     1501 tccatgtgtg aaaagatgag aatacgtatt ttagattggg atcgtgcgag tcataatgat
     1561 gtcattggca ctgcacactt atgcatgtct aaaatctcag cccctggagg agatattgaa
     1621 gtggatgaca gtataggctt tcttccgacc tttggaccct gctttgtcaa catgtatggg
     1681 agtcctcgag agttcacagc tttctcagac ccacacgagg cactaaacct gggaaaggga
     1741 gagggtgtgg cctatcgggg acgtgtgttg ttggagctca taacaaagct ttctgatcag
     1801 gtggaccaaa ggtctgaaga catcaactct gatgacctgc tagtggtaga aaaattcatg
     1861 cggaagagga agttttccct gtttgctgcg ttctactcgg ccactctcct tcaagatgtg
     1921 gacgatgcag ttcagtttga agtcagtatc ggtaactacg gcaacaagtt cgacaacacc
     1981 tgtttgcctc tggcatccac cactcaattc agcagagctg tgttcgacgg ctgccattac
     2041 tattacctgc catggggaaa tgtgaaacct gtggtggtgg tgtcttcaga gtgggaagac
     2101 attggccctc gtatagaggc cctcaacatg ctgcttagca tagtcgaccg actggagtca
     2161 aacttggaac aggtacagct ggagctcaaa gcaagaagtg acggtcagga agtcgagcag
     2221 agggttgtcc agctgcttga tgatgtcatt gcagactgca gtgaggaact ccctgagttg
     2281 gacaaatggc cttgtgtgac tccactggat aaagctctga ggaatctgcg caagattaac
     2341 atgcagcaga tagtgcaggc ggccctggca ctcaaacatg gaccagagac tgaactgagt
     2401 gcagtgctgg agcaggctga ggactggacc atgagactac gctacctctc tgaggagccc
     2461 caaaacagtc tgccagatat aataatctgg atgcttcagg gagacagacg agtggcctac
     2521 cacagaatcc ctgcccatca agttctcttt tcctatggtt actgtggcaa gcactgtggc
     2581 caacttcaga cggttttcat gaagtgtcca caggaagatg ggggatgtag taaaatccca
     2641 gctcagctca gaatcaaggt gtggttcgga ctgactgcgg atgtcaaatc ttttaaccag
     2701 tttgcagagg gcaaactctc agtgtttgca gaaacgtatg agaaccagac taaactggct
     2761 ctggtgggga actggggtac gacgggtctg acttacccca agttcagtga tgtgaccggc
     2821 agagtcaaac tgccaaagga gagcttcaaa ccctcttcag gctggagctg ggcaggagaa
     2881 tggttcatca gcccagagaa aacgatgctg tatgatgctg atgcaggtca tataagcttt
     2941 ttggaggagg tgtttgaaaa tcagttgcgg ttgccaggag gacagtggat tggcatgccg
     3001 gaggggtaca cagatgtgaa tggagaaaag gctgttccta aagatgagat cgagtgtccc
     3061 cctggctggg tgtgggagga tattgagtgg agcgaggatc tgaagcgtgc agtggatgat
     3121 caagggtggg aatatggaat aaccctcccc ccggaccgcc gaccaaagtc ctgggttcca
     3181 tctgagaaga tgtaccatac caaccgtcgg agacgctgga tcagactccg ccgtagagac
     3241 atgcagaaaa tggaggcgct cagaaaacag cggccagacg agtccgagcg tgagggctgg
     3301 gaatatgcct ctttatttgg ctggaagttt cacctcaagc agaagaaaac cgacttgttc
     3361 cgtcgccggc gctggagacg tcgaatggag ccattagaaa agacagggcc ggcggccatc
     3421 tttgccttgg agtgttcgct gagcagcttt actgatgaaa agaatgacga taagtcagtc
     3481 actacaacgt ttggtgtcag ccggccgact atctcctgct tctttgatag tgggaccctg
     3541 tatcatttgc gctgttactt gtatcaggcc agagacctgc tggctatgga caaggacagt
     3601 ttctcggatg cctatgccgt tgtatcgttc ctgcatcaga gtcagaagac ggtgactgtt
     3661 cgaaacacac tcaaccccac ctgggaccag accctcattt tctacgaagt ggaaattttt
     3721 ggagaccatc ttgtgactga gaggagtcct ccaaacattg tggtggagct gtacgatcag
     3781 gacacctatg gagctgatga gttcatgggc cgctgtgttt gccaaccatc aatggcatct
     3841 tcccctcgcc tcgcctggta tcccatccag aaagcaggga attctgcagg agatctgctt
     3901 gctgcttttg aactgatccg cagagataaa cctgcagttc atcacatccc gggtcaagag
     3961 ggagatatta taccaccctc acatatcatg gatgagcttg tgcttccagg gttcctctct
     4021 gagaatcaac agcaatggcc agaggactct gatttgccct atccaccacc tcagagggag
     4081 ccaaacgtct acatggtgcc acaggggata aaacctgtcc tacagcgcac tgctattgag
     4141 atactggcat ggggtttgcg taacctgaag agtttccagc tggccagtgt cacgtctccc
     4201 agtcttcagg tggaatgtgg cggatctgtg gttcagtcgt gcgtcattcg aaatattaag
     4261 aagaacccta attttgacgt caacatcctg tccattgatg tccgattgcc taaggaagag
     4321 ctctacatgc ccccattggt gatcaaggtt atagacaacc ggcagtttgg caggaagcct
     4381 gtggtgggcc agtgttcagt ccactcgctc aatgagttct acttccagcc ggaaagtctg
     4441 gagagagact caacagaaga ggaatttgaa gttattgctc acacacctag agatgatgtt
     4501 ctggttgata ttgatgacaa ggagcctcta atccctggac agcaggatga atttatggac
     4561 tggtggagca agttttatgc ctccatgggt gaaaggacca agtgtgggag ttatctggag
     4621 aaagggttcg atatgctgca ggtgtatgac agagaactgg agaagactga aggctttgag
     4681 caatttgctg atttctgtca aacgtttaaa ctttaccgtg gaagaaccca gaatgacagc
     4741 gaagatcctt ccgttgttgg agagttcaag ggttcgttta agatctatcc attgcctgat
     4801 gatcccccta tgcctgtacc cacacgtcag tttcgcaaac tccccccgaa cggtatagag
     4861 gagtgcttgg tacgcatcta catcatccag gcttttgggt tacagcccaa agatgccaat
     4921 ggcaagtgtg acccctacgt taagatcacg cttggcaaga aatcagtcga tgaccacgac
     4981 aattacatac cctgtacttt ggagcccgtc tttggaaaga tgtttgagct gacctgctgt
     5041 ctgcctcttg agaaagacct gaagattaca ctctatgatt acgacatggt gactaaagat
     5101 gagaaaatcg gagagaccat cattgatttg gagaaccgct tcttgtctcg tcatggtgct
     5161 cgctgcggcc ttccgcagtc ctactgtctg tcaggaatta accagtggcg agatcaactt
     5221 aaaccatctc agctacttca tcgtctgtgt gagaggcgaa actacaaact ccccgtctac
     5281 aagcaggatc gaatttactt cagaggacat gaatacactg tcgcagacct tgatgaaggg
     5341 aaacctctta accctcactt gggaccagtg atggagcgtc tggctctgtt ggtgttgagg
     5401 aggcatggtt tggtgcctga acatgtcgag acgcgacccc tctacagccc ccttcagcca
     5461 gatatagaac aggggcgact acaaatgtgg gtggacctgt tccctaagtc tttaggccct
     5521 ccaggtcctc cattcaacat caccccacgc aaagctaaaa agttcttcct gcgctgtatt
     5581 atatggaaca caagagatgt aatcttggat gatgtcagcg ttaccgggga gaagatgagt
     5641 gacatttatg tgaaaggctg gatgcacggc catgaggaga acaagcagaa aacagacgtg
     5701 cactaccgat cgctgggtgg agagggtaac tttaactggc gcttcctctt ccccttccac
     5761 tatttacctg cggaacaact gtgcaccatt gataaaaagg agcacttttg gagtctggat
     5821 aaagttgaaa cgaagcttcc tcctaaactt actatccaga tttgggacaa tgacaagttc
     5881 tcctttgatg atttcctcgg ctctttggag atggatctga accacatgct gagtcctgcg
     5941 aagtctccag agaagtgtgg tcttgagatg ctcactcagt cccaggacaa actggtctct
     6001 ttgcttgagc agaaaacggt gaaaggctgg tggccgtgtg tctgtgacaa agaagacaag
     6061 aagatacttg ctggaaaggt ggagatgtct ctggaaatag tctgtgaaca ggagcaggat
     6121 gagaggcctg caggactcgg ccgagatgag cccaacatga atccccatct agaggagcct
     6181 cagcgtcccg agacatcatt cttatggttc tcgtccccat ataagacgtt taggttcatt
     6241 gtgtggagga ggtttaagtg gcttattctg ggcctcttta tactcctttt cattattctc
     6301 ttccttggag tatttctcta cgccttccct aactatgctg ctatgaagat ggttggaccc
     6361 ttcagtggac taggaaaggg aacacagtga caactttatt atacaaagtt ggcattataa
     6421 aaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     6481 ttatttggag ctccatggta gcgttaacgc ggccgcgata tcccctatag tgagtcgtat
     6541 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     6601 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     6661 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     6721 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     6781 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     6841 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     6901 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     6961 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     7021 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     7081 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     7141 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     7201 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     7261 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     7321 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     7381 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     7441 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     7501 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     7561 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     7621 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7681 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7741 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7801 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7861 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7921 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7981 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8041 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8101 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8161 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8221 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8281 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8341 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8401 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     8461 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     8521 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     8581 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     8641 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     8701 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     8761 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     8821 aaaacgacgg ccagtcttaa gctcgggccc tgcagctcta gagctcgaat tctacaggtc
     8881 actaatacca tctaagtagt tggtt
//
