LOCUS       sequence-249608-        3161 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 3161)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 3161)
  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-028   Sirius(G-H)//Esp3I
FEATURES             Location/Qualifiers
     source          1..3161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    779..795
                     /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(803..809,810..827,828..833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    848..869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(986..1003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1157..1745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1237..1256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2054..2848
                     /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(2108..2127)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2718..2737
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2922..2940
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2978..3000)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3100..3119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcgcag cgtgagcaaa ggagaggagc tctttacagg agtagtcccg atattggtgg
       61 aacttgatgg cgacgtcaat ggacaccgtt tctccgtgag tggagaaggc gaaggcgatg
      121 caacctatgg taaactcacc ctgaagctta tctgcactac ggggaaactg ccagtacctt
      181 ggccaaccct ggttacaacc ctgcagtttg gggttctgtg ttttgcccga tatcccgatc
      241 acatgaagca gcatgacttc ttcaagtcag ccatgccaga agggtatgtc caagagagga
      301 ccatcttctt taaagacgat ggcaactata agacgagagc agaagtgaag tttgagggag
      361 acactctggt gaatcggatt gaactcaaag gcatcgactt caaggaagat ggcaatatcc
      421 tgggtcacaa actggagtac aatgggatct ctagcaacgt gtacatcaca gccgataagc
      481 agaagaacgg gataaaggcg catttcaaga ttcgccataa catcgaggac ggtggcgttc
      541 aactcgctga tcactaccag cagaacactc ccattggcga tggacctgtg cttctccctg
      601 acaaccacta cttgtctgtc cagagcaagc tgtccaaaga ccccaatgag aagagggacc
      661 atatggtctt gctggagagt gtgacagctg ctgggattac tctgggtatg gacgagctgt
      721 acaaatcagg ccgagacgac ctgctagcgc tcgcttggat ccgaattcaa acctgaaatt
      781 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      841 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      901 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      961 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1021 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1081 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1141 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1201 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1261 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1321 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1381 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1441 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1501 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1561 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1621 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1681 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1741 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1801 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1861 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagccgga
     1921 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     1981 tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg
     2041 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2101 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2161 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2221 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2281 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2341 agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat
     2401 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2461 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2521 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2581 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2641 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     2701 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     2761 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     2821 ctatcgcctt cttgacgagt tcttctgatc cgcgcacatt tccccgaaaa gtgccacctg
     2881 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2941 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3001 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3061 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     3121 tgagagaaac ccaattgggt accgagctcc ctgcaggaag c
//
