LOCUS       sequence-271316-        3314 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 3314)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 3314)
  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-034   EB3(D-G)//Esp3I
FEATURES             Location/Qualifiers
     source          1..3314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      12..21
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    932..948
                     /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(956..962,963..980,981..986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1001..1022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1139..1156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1310..1898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1390..1409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2207..3001
                     /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(2261..2280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2871..2890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3075..3093
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3131..3153)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3253..3272
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcggga cgccaccatg gccgtgaacg tgtactcaac cagcgtaaca agcgagaatc
       61 tgtcaaggca cgacatgttg gcttgggtga acgattccct tcatctgaac tacaccaaaa
      121 tcgaacagct gtgctctgga gctgcctatt gccagtttat ggacatgctc tttcccggat
      181 gtgtgcacct tcggaaagtc aaattccagg caaaactgga acatgagtat atccacaact
      241 ttaaggttct gcaagcagct tttaagaaga tgggcgtgga caagatcatc ccggtggaaa
      301 agctcgtgaa agggaagttt caggacaatt tcgagttcat tcagtggttc aagaagttct
      361 ttgacgccaa ctatgatggg aaagattaca atccactgct tgccagacag ggacaggatg
      421 tcgcaccgcc acccaatcct ggcgaccaga tcttcaacaa gagcaagaaa ctgattggga
      481 cagcggttcc acaacgtact tcccctactg gccccaagaa catgcagacg tctggcagac
      541 tgagcaatgt tgctccaccc tgtatactcc gcaagaatcc acctagtgct cgaaatggcg
      601 gtcatgagac agatgcccag atactggagc tcaaccaaca gctggtagac ctgaaactga
      661 ccgtcgacgg attggagaaa gagagggact tctacttctc caagttgcgg gatatagagc
      721 tcatttgcca ggaacacgag agcgaaaact ctcccgtcat cagtggtatc atcggtattc
      781 tgtatgccac tgaagaaggg tttgcacctc ctgaagatga cgaaattgag gagcaccaac
      841 aggaggatca agacgagtac ctggatggcg ggtcaggcgg cagccgagac gacctgctag
      901 cgctcgcttg gatccgaatt caaacctgaa attgttatcc gctcacaatt ccacacaaca
      961 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1021 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1081 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1741 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1921 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1981 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2041 gtatatatga gtaaacttgg tctgacagcc ggaattgcca gctggggcgc cctctggtaa
     2101 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     2161 caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2221 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2281 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2341 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2401 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2461 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2521 ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2581 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2641 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2701 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     2761 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2821 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2881 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     2941 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3001 atccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     3061 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     3121 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     3181 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     3241 ggcttaacta tgcggcatca gagcagattg tactgagaga aacccaattg ggtaccgagc
     3301 tccctgcagg aagc
//
