LOCUS       sequence-222117-        3265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with Clathrin light chain b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3265)
  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 3265)
  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-Cltb
FEATURES             Location/Qualifiers
     source          1..3265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..25)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(651..746)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(783..802)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(784..802)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(807..823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(807..823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(925..944)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             936..1745
                     /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(1003..1022)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1892..2480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2381..2400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2634..2651
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2810..2837)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2929..3015)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3064..3086
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3078..3095
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3079..3095
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggtggctg acgacttcgg tttttccgcc ttcgacaacg
       61 gagttcatac agaagaagat cccgccgccg cctttctggc ccagcaggag aatgaaatcg
      121 cggtgattga gaacgacagc actgggttca gtttaccgga ggacggagga gcggcgcccg
      181 cagacaccta caccatgtcc ggcggtgact ttggaggatc ctcagctgta aacggagaca
      241 tgtttcagga cacgaacggc atgactgaca gctactcggc tatcgcacaa gtcgacatac
      301 agagacaaga gcctgaaagt ctgcgcaagt ggagggagga gcagaagacc cggctggagg
      361 aattagattc agcctctaaa gctgcagagg ccgtgtggag ggagaaggcc aagaaggagc
      421 tggaggactg gcatatacac cagactgaac agatggagaa gaataaagtc aacaacagag
      481 cgtcagagga ggctttcctc aaggagtgtg aagatgacag tcctgggacg gaatgggaga
      541 aagtggcccg actctgcgat ttcaacccca aaaccagccg gcagactaaa gacgtgtccc
      601 gaatgcgctc ggtgctcatt tccctgaaac agacgcctct gctccgctaa caactttatt
      661 atacaaagtt ggcattataa aaaagcattg cttatcaatt tgttgcaacg aacaggtcac
      721 tatcagtcaa aataaaatca ttatttggag ctccatggta gcgttaacgc ggccgcgata
      781 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
      841 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
      901 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
      961 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1021 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     1081 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1141 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1201 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     1261 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     1321 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     1381 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     1441 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     1501 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     1561 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     1621 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     1681 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     1741 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     1801 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     1861 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1921 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1981 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2041 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2101 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2161 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2221 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2281 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2341 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2401 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2461 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2521 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2581 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2641 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2701 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2761 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     2821 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     2881 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     2941 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     3001 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
     3061 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc tgcagctcta
     3121 gagctcgaat tctacaggtc actaatacca tctaagtagt tggttcatag tgactgcata
     3181 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
     3241 atatttatat cattttacgt ttctc
//
