LOCUS       sequence-223873-        3310 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Multisite gateway vector for 5' tagging with lamp1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3310)
  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 3310)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-lamp1
FEATURES             Location/Qualifiers
     source          1..3310
                     /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(867..966)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(983..1002)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(984..1002)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1007..1023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1007..1023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1125..1144)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1136..1945
                     /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(1203..1222)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2092..2680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2581..2600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2834..2851
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3010..3037)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3129..3215)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3264..3286
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3278..3295
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3279..3295
                     /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 gccaccatga tctctatcat
      121 ccccaaacaa aacagacccg tcggggcaat ttttagcctg ctgttagctc taaccataca
      181 ccagactttg acctccaatg ttccgaccac tgcggccacc cctgaaacca gctctcctcc
      241 ggcctctcct gtcacccccg gacccccaga gcggggcaac tacaatgtca ccaatgccaa
      301 tgatacagtc tgtctcctgg cacgcatggc cctccaactc aacataagct tcaactcaac
      361 ctctcagggc aagcttgtcc aggaggtctt gaacttgcat ccaaacctcg ttaaattctc
      421 tggatcctgt gagttggacc gtgcatctct gattctcaca gaggacaaca ttactctgac
      481 tttcagcttc tctttaaact ccacatcaaa caagtatcat cttagtggtt tggagctgtc
      541 agcttcattg cctgatatgg ctaagggtct tgtggtcaga aacaccacac tgaactactt
      601 tgtgggaacg ctgggccact cttacatgtg tcagaaggaa aagaccttga gcgtcaccca
      661 aaatttctct ctcaacacct tccaacttca agtgcaacct ttcggtgtca agggagattt
      721 tggagcagct gaggagtgcg agttggatga ggacgacatg ttgatcccca tcgttgtggg
      781 cgctgcattg gctgttctag tgctcatagt gatcctggcg tacctgatcg gcagaaacag
      841 gagtcacgcc ggctaccaga ccatccaccc agctttcttg tacaaagttg gcattataag
      901 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
      961 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1021 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1081 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1141 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1201 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     1261 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1321 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     1381 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     1441 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     1501 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     1561 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     1621 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     1681 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     1741 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     1801 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     1861 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     1921 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     1981 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2041 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2101 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2161 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2221 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2281 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2341 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2401 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2461 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2521 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2581 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2641 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2701 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2761 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2821 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     2881 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     2941 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
     3001 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
     3061 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
     3121 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     3181 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
     3241 gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
     3301 gctcgggccc
//
