LOCUS       sequence-325150-        3244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway middle entry vector for fusing mScarlet to the 5' 
            end of an ORF..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3244)
  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 3244)
  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-NS
FEATURES             Location/Qualifiers
     source          1..3244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     217..234
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(393..420)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(512..598)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     647..669
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     661..678
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     662..678
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    694..793
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      794..803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1494..1593)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1610..1629)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1611..1629)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1634..1650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1634..1650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1752..1771)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1763..2572
                     /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(1830..1849)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(2719..3244,1..63)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3208..3227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
       61 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      241 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
      301 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
      361 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
      421 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
      481 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
      541 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
      601 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
      661 tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata
      721 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac
      781 aaaaaagcag gctgccacca tggtatctaa aggagaggct gtcatcaaag agtttatgag
      841 atttaaagtc cacatggagg gaagcatgaa cggtcatgag tttgaaatag aaggagaagg
      901 tgagggtcga ccttacgaag gtacccagac cgcgaagttg aaagttacca aagggggccc
      961 cttgccgttc agttgggaca tcctgtcccc tcaatttatg tacggaagca gggcttttac
     1021 gaaacatccg gctgacattc ccgattacta taagcaaagt ttccccgagg gctttaagtg
     1081 ggagcgcgta atgaattttg aggatggcgg tgctgttaca gtcacacagg atacatccct
     1141 cgaagacgga acgttgatct acaaagttaa acttcgcggg actaacttcc cgcccgatgg
     1201 ccctgtgatg caaaagaaga ctatgggatg ggaggccagc acggaaagac tctaccctga
     1261 agatggcgta cttaaaggtg atataaaaat ggctctgcgc ctgaaggatg gaggtcgtta
     1321 cctcgccgat ttcaagacca cctacaaagc caaaaaaccc gtgcagatgc ctggcgcata
     1381 taatgtagat aggaagctgg acatcactag ccataatgaa gattacaccg ttgtagagca
     1441 atacgaacga tcagagggcc gtcactcaac cggcggtatg gatgaactct ataagccagc
     1501 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1561 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1621 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1681 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1741 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1801 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1861 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1921 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1981 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2041 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2101 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2161 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2221 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2281 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2341 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2401 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2461 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2521 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2581 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2641 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2701 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2761 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2821 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2881 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2941 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3001 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3061 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3121 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3181 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3241 gttt
//
