LOCUS       sequence-325282-        3337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway middle entry vector for fusing mScarlet to the 3' 
            end of an ORF..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3337)
  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 3337)
  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-mScarlet
FEATURES             Location/Qualifiers
     source          1..3337
                     /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(1446..1541)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1578..1597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1579..1597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1602..1618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1602..1618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1720..1739)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1731..2540
                     /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(1798..1817)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2687..3275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3176..3195
                     /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 aaagtggtgg tatctaaagg agaggctgtc atcaaagagt
      781 ttatgagatt taaagtccac atggagggaa gcatgaacgg tcatgagttt gaaatagaag
      841 gagaaggtga gggtcgacct tacgaaggta cccagaccgc gaagttgaaa gttaccaaag
      901 ggggcccctt gccgttcagt tgggacatcc tgtcccctca atttatgtac ggaagcaggg
      961 cttttacgaa acatccggct gacattcccg attactataa gcaaagtttc cccgagggct
     1021 ttaagtggga gcgcgtaatg aattttgagg atggcggtgc tgttacagtc acacaggata
     1081 catccctcga agacggaacg ttgatctaca aagttaaact tcgcgggact aacttcccgc
     1141 ccgatggccc tgtgatgcaa aagaagacta tgggatggga ggccagcacg gaaagactct
     1201 accctgaaga tggcgtactt aaaggtgata taaaaatggc tctgcgcctg aaggatggag
     1261 gtcgttacct cgccgatttc aagaccacct acaaagccaa aaaacccgtg cagatgcctg
     1321 gcgcatataa tgtagatagg aagctggaca tcactagcca taatgaagat tacaccgttg
     1381 tagagcaata cgaacgatca gagggccgtc actcaaccgg cggtatggat gaactctata
     1441 agtaacaact ttattataca aagttggcat tataaaaaag cattgcttat caatttgttg
     1501 caacgaacag gtcactatca gtcaaaataa aatcattatt tggagctcca tggtagcgtt
     1561 aacgcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     1621 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     1681 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     1741 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     1801 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     1861 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     1921 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     1981 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     2041 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2101 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2161 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2221 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2281 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2341 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2401 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2461 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2521 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     2581 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     2641 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2701 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2761 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2821 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2881 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2941 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3001 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3061 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3121 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3181 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3241 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3301 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
