LOCUS       sequence-274126-        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     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(1369..1468)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1485..1504)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1486..1504)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1509..1525)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1509..1525)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1627..1646)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1638..2447
                     /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(1705..1724)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2594..3182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3083..3102
                     /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 ccagctttct
     1381 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     1441 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt
     1501 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     1561 acattgcaca agataaaata atatcatcat gaacaataaa actgtctgct tacataaaca
     1621 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     1681 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1741 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1801 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     1861 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1921 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     1981 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     2041 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     2101 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     2161 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     2221 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2281 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     2341 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     2401 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     2461 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     2521 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     2581 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2641 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2701 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2761 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2821 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2881 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2941 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3001 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3061 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3121 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3181 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3241 tgtt
//
