LOCUS       sequence-117201-        3109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway entry clone for adding C-terminal fusions of 
            ML1N*2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3109)
  AUTHORS   Ariotti N, Hall TE, Rae J, Ferguson C, McMahon KA, Martel N, Webb 
            RE, Webb RI, Teasdale RD, Parton RG
  TITLE     Modular Detection of GFP-Labeled Proteins for Rapid Screening by 
            Electron Microscopy in Cells and Organisms.
  JOURNAL   Dev Cell. 2015 Nov 23;35(4):513-25. doi: 
            10.1016/j.devcel.2015.10.016. Epub 2015 Nov 12.
  PUBMED    26585296
REFERENCE   2  (bases 1 to 3109)
  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"; doi: 
            "10.1016/j.devcel.2015.10.016"; journalName: "Dev Cell"; date: 
            "2015-11-23- 23"; volume: "35"; issue: "4"; pages: "513-25"
COMMENT     Sequence Label: p3E-ML1N*2
FEATURES             Location/Qualifiers
     source          1..3109
                     /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..748)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1218..1313)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1350..1369)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1351..1369)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1374..1390)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1374..1390)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1492..1511)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1503..2312
                     /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(1570..1589)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2459..3047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2948..2967
                     /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 aaagtggtgt actctgacct gactatggca acacctgctg
      781 gacggagggc aagcgagacc gaacggctgc tgacacccaa ccccggatac ggcactcaag
      841 tgggcacatc tccagctccc accacaccca ctgaggaaga ggacctgagg agacggctga
      901 agtacttctt tatgagtcca tgcgataaat tccgcgcaaa ggggcgaaaa ccctgtaagc
      961 tgatgctgag aatcctgaca atggcaactc cagctggacg acgagcaagc gagactgaaa
     1021 gactgctgac ccctaaccca ggctacggaa cccaggtcgg aacatcccca gcacctacta
     1081 cccctaccga agaggaagac ctgagaaggc ggctgaagta tttctttatg agcccttgcg
     1141 ataaatttcg ggccaaaggg cgcaagccat gtaaactgat gctggacccc cctgacctgg
     1201 ataattaagg atccgaacaa ctttattata caaagttggc attataaaaa agcattgctt
     1261 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggagctc
     1321 catggtagcg ttaacgcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     1381 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     1441 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1501 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     1561 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     1621 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     1681 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     1741 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     1801 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1861 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1921 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1981 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     2041 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2101 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2161 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     2221 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     2281 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     2341 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     2401 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2461 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2521 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2581 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2641 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2701 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2761 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2821 acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga
     2881 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2941 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3001 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3061 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
