LOCUS       sequence-206479-        3394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for expression of mKate2 fused to the 5' 
            and 3' sequences from zebrafish folate receptor 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3394)
  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 3394)
  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-mKate2-GPI-pA
FEATURES             Location/Qualifiers
     source          1..3394
                     /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"
     CDS             741..1433
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     protein_bind    complement(1519..1618)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1635..1654)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1636..1654)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1659..1675)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1659..1675)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1777..1796)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1788..2597
                     /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(1855..1874)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2744..3332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3233..3252
                     /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 caccatggac gctttatttc acagacgtct gctgctgctg ctgattgtac
      721 tgccggcctg cagcctcgag gtgagcgagc tgattaagga gaacatgcac atgaagctgt
      781 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
      841 cctacgaggg cacccagacc atgagaatca aggcggtcga gggcggccct ctccccttcg
      901 ccttcgacat cctggctacc agcttcatgt acggcagcaa aaccttcatc aaccacaccc
      961 agggcatccc cgacttcttt aagcagtcct tccccgaggg cttcacatgg gagagagtca
     1021 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     1081 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
     1141 agaagaaaac actcggctgg gaggcctcca ccgagaccct gtaccccgct gacggcggcc
     1201 tggaaggcag agccgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
     1261 tgaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     1321 atgtggacag aagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     1381 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac agaggatccc
     1441 tgaagggcgg tacggagacg gtcgccatgg caacaggtct gcatctgatg gcgctgctgg
     1501 tgatgctgct gcgctgacac ccagctttct tgtacaaagt tggcattata agaaagcatt
     1561 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     1621 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     1681 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaata atatcatcat
     1741 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     1801 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     1861 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     1921 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     1981 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2041 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2101 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2161 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2221 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2281 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     2341 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     2401 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     2461 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     2521 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     2581 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     2641 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     2701 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2761 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2821 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2881 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2941 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3001 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3061 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3121 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3181 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3241 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3301 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3361 tcctggcctt ttgctggcct tttgctcaca tgtt
//
