LOCUS       sequence-325152-        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     33..52
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     286..303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(462..489)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(581..667)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     716..738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     730..747
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     731..747
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    763..862
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      863..872
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1567..1666)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1683..1702)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1684..1702)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1707..1723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1707..1723)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1825..1844)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1836..2645
                     /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(1903..1922)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(2792..3248,1..132)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
       61 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      301 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
      361 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      421 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
      481 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
      541 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
      601 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
      661 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
      721 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt
      781 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
      841 actttgtaca aaaaagcagg ctgccaccat ggtatctaaa ggagaggctg tcatcaaaga
      901 gtttatgaga tttaaagtcc acatggaggg aagcatgaac ggtcatgagt ttgaaataga
      961 aggagaaggt gagggtcgac cttacgaagg tacccagacc gcgaagttga aagttaccaa
     1021 agggggcccc ttgccgttca gttgggacat cctgtcccct caatttatgt acggaagcag
     1081 ggcttttacg aaacatccgg ctgacattcc cgattactat aagcaaagtt tccccgaggg
     1141 ctttaagtgg gagcgcgtaa tgaattttga ggatggcggt gctgttacag tcacacagga
     1201 tacatccctc gaagacggaa cgttgatcta caaagttaaa cttcgcggga ctaacttccc
     1261 gcccgatggc cctgtgatgc aaaagaagac tatgggatgg gaggccagca cggaaagact
     1321 ctaccctgaa gatggcgtac ttaaaggtga tataaaaatg gctctgcgcc tgaaggatgg
     1381 aggtcgttac ctcgccgatt tcaagaccac ctacaaagcc aaaaaacccg tgcagatgcc
     1441 tggcgcatat aatgtagata ggaagctgga catcactagc cataatgaag attacaccgt
     1501 tgtagagcaa tacgaacgat cagagggccg tcactcaacc ggcggtatgg atgaactcta
     1561 taagtaaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     1621 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     1681 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     1741 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1801 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     1861 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1921 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     1981 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     2041 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2101 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     2161 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2221 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2281 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2341 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2401 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2461 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2521 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2581 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2641 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     2701 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     2761 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2821 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2881 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2941 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3001 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3061 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3121 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3181 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3241 gcagggtc
//
