LOCUS       sequence-223877-        3727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 5' tagging with CD44a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3727)
  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 3727)
  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-CD44a
FEATURES             Location/Qualifiers
     source          1..3727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1284..1383)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1400..1419)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1401..1419)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1424..1440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1424..1440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1542..1561)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1553..2362
                     /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(1620..1639)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2509..3097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2998..3017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3251..3268
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3427..3454)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3546..3632)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3681..3703
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3695..3712
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3696..3712
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct gccaccatgt ggactttatt
      121 atttgtagtg tttgctactg gatcaccagc tgtacttcag gcagccccgg ttcagggtaa
      181 cagtgggaga tgcagttttg ctggtgtttt ccacatcgag ggaatcagtc gttactctct
      241 tacctttcag aaggctggcg aactgtgtca gagtttaggt tacagacttg caacacaaga
      301 acaggtcact gaagcctata aaaaaggctt gagaacttgc agatacggct ggattgatgg
      361 tcaaaatgtc acgttcctcc cacacattaa tggtccaaac tgtgactcca gctctgctga
      421 gatcaccttc catgctaaag caggagaatt tctttctgat gtctactgtt ttgacccatc
      481 agattcatca ccaaactgtg atgatgttaa atctgaggat gttatgaatg gaaacgcagc
      541 cagagacagt tcattctatg caaaaccaca tgaagacttc attacagaat tcaagacaga
      601 agggtttttg gtagatgtgg aagaaatatt ggaaagagta aaaagagaaa catctcttct
      661 gagtgacatg aaaccctcaa cagtgccctc tatgaaaaaa attgaaggca tcaaggaccc
      721 tacaagcaag cctcatattg gaaagacccc cagtgttttg ctatttgata cagagggcag
      781 tggatcggga ttggttgatc ctgagcctag ccatttcaca acaagtccgg ttactacaga
      841 gacagcgaag agtgatttaa ttgaaaacgt tattgaagag aatgaagaga tggtggttcc
      901 agtagagact tcaaatgtga accccattca agtcaaggaa ccaagacgag tgaatgtatt
      961 ttcaacaaca gaaagtaatg cgaaggagat ggtttcagat ggatcgtctt ctacttggat
     1021 gataattact gcattctgtg tgtttattgg agtcatagta tgcatatttg tggctattgg
     1081 cactagagac aagtggtacg gaccgagtaa gagtgcagac atcacaactg aaaagaacaa
     1141 caaagactac agtaaaacag aaacgctgcc gctgtcagag aaagagcagg agattgtggc
     1201 actgatgaat gttacgaaca tgaaaaatgg gcaaacagtg gacataacag ctgcagatga
     1261 acacgaaaaa gaatatttaa tgcacccagc tttcttgtac aaagttggca ttataagaaa
     1321 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1381 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     1441 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1501 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     1561 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     1621 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     1681 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1741 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1801 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     1861 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     1921 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     1981 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2041 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2101 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2161 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2221 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2281 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2341 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     2401 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     2461 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2641 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3121 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3181 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3241 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3301 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3361 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     3421 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     3481 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     3541 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     3601 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     3661 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
     3721 cgggccc
//
