LOCUS       sequence-249617-        2537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA Kit # 1000000119 extension.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2537)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 2537)
  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"
COMMENT     Sequence Label: pRDAX-EMMI-037   NLSNoLS(D-G)//Esp3I
FEATURES             Location/Qualifiers
     source          1..2537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             21..68
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    155..171
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(179..185,186..203,204..209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    224..245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(362..379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(533..1121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(613..632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1430..2224
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1484..1503)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2094..2113
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2298..2316
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2354..2376)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2476..2495
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcggga cgccaccatg aaacggcctg ctgccaccaa gaaagcagga caggccaaga
       61 agaagaaacg caggagaagg cgacgtagag gcggtagcgg gggcagccga gacgacctgc
      121 tagcgctcgc ttggatccga attcaaacct gaaattgtta tccgctcaca attccacaca
      181 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      241 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      961 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1141 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1201 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1261 aaagtatata tgagtaaact tggtctgaca gccggaattg ccagctgggg cgccctctgg
     1321 taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg
     1381 gcgcagggga tcaagatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     1441 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     1501 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     1561 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     1621 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     1681 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     1741 atcccacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     1801 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     1861 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     1921 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     1981 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     2041 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     2101 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     2161 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     2221 ctgatccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     2281 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2341 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2401 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2461 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agaaacccaa ttgggtaccg
     2521 agctccctgc aggaagc
//
