LOCUS       sequence-274124-        3248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway middle entry vector for expression of untagged 
            mScarlet..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3248)
  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 3248)
  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: pME-mScarlet-STOP
FEATURES             Location/Qualifiers
     source          1..3248
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1373..1472)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1489..1508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1490..1508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1513..1529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1513..1529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1631..1650)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1642..2451
                     /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(1709..1728)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2598..3186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3087..3106
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc caccatggta tctaaaggag aggctgtcat caaagagttt atgagattta
      721 aagtccacat ggagggaagc atgaacggtc atgagtttga aatagaagga gaaggtgagg
      781 gtcgacctta cgaaggtacc cagaccgcga agttgaaagt taccaaaggg ggccccttgc
      841 cgttcagttg ggacatcctg tcccctcaat ttatgtacgg aagcagggct tttacgaaac
      901 atccggctga cattcccgat tactataagc aaagtttccc cgagggcttt aagtgggagc
      961 gcgtaatgaa ttttgaggat ggcggtgctg ttacagtcac acaggataca tccctcgaag
     1021 acggaacgtt gatctacaaa gttaaacttc gcgggactaa cttcccgccc gatggccctg
     1081 tgatgcaaaa gaagactatg ggatgggagg ccagcacgga aagactctac cctgaagatg
     1141 gcgtacttaa aggtgatata aaaatggctc tgcgcctgaa ggatggaggt cgttacctcg
     1201 ccgatttcaa gaccacctac aaagccaaaa aacccgtgca gatgcctggc gcatataatg
     1261 tagataggaa gctggacatc actagccata atgaagatta caccgttgta gagcaatacg
     1321 aacgatcaga gggccgtcac tcaaccggcg gtatggatga actctataag taacccagct
     1381 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1441 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1501 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1561 tgttacattg cacaagataa aataatatca tcatgaacaa taaaactgtc tgcttacata
     1621 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1681 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1741 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1801 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1861 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1921 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1981 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2041 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2101 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2161 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     2221 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2281 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2341 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2401 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2461 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2521 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3241 cacatgtt
//
