LOCUS       sequence-222017-        3118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with the C-terminal domain 
            from cow (Bos taurus) milk fat globue EGF-factor 8 protein (MFGE8). 
            Binds phosphatidylserine (PS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3118)
  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 3118)
  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-Lact-C2
FEATURES             Location/Qualifiers
     source          1..3118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..96
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    576..699
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(770..787)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(770..786)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(779..801)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      850..936
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1028..1055
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1214..1231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1385..1973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1465..1484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2120..2929)
                     /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     2843..2862
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2921..2940
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3042..3058
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3042..3058
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3063..3082
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3063..3081
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat
       61 gcttttttat aatgccaact ttgtataata aagttgctaa cagcccagca gctccactcg
      121 cagggtgata cggttgtgcc aggccacggg ctggatccgc acgaagcgag cctggaacgg
      181 cgtctcaaat atgttcttct tgtgggaatt attgtccatg ttaccaggga aaatcttgct
      241 ttctgaggcc cccgggtcct tgtactcagt ccaggtcaca ccatcatcac cataggccac
      301 cctgtaggca gccacatatt gaatgtggcc aaagtctcgg gcaccctggg tgatgatgcc
      361 cgtgactcgc ttctgggagc ccaggtcaat ctgcagccac tcagaggcac tgttggtctg
      421 ggcggtccag gcgttgaact tgccctgatt atccagtcgt gcgtagtagg gaaaccagct
      481 aaaggcactc aggccccagg ttttgtagta gctggaggct gtgatctgct tgttggggat
      541 ggtattatcc ttcaggccta ggggttcagt gcaagccact ttgtacaaga aagttgaacg
      601 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac
      661 tacataatac tgtaaaacac aacatatgca gtcactatga accaactact tagatggtat
      721 tagtgacctg tagaattcga gctctagagc tgcagggccc gagcttaaga ctggccgtcg
      781 ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt taacgcgaga gtagggaact
      841 gccaggcatc aaataaaacg aaaggctcag tcggaagact gggcctttcg ttttatctgt
      901 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
      961 gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     1021 actaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttcctggc
     1081 tagcggtacg cgtattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1141 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1201 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1261 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1321 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1381 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1441 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1501 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1561 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1621 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1681 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1741 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1801 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1861 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1921 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1981 ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgtaactc acgttaaggg
     2041 attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca
     2101 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     2161 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     2221 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     2281 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     2341 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     2401 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     2461 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     2521 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     2581 tttcacctga atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag
     2641 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     2701 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     2761 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg
     2821 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     2881 tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg
     2941 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     3001 caatgtaaca tcagagattt tgagacacgg gccagagctg ccaggaaaca gctatgacca
     3061 tgtaatacga ctcactatag gggatatcgc ggccgcgtta acgctaccat ggagctcc
//
