LOCUS       sequence-249611-        3122 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 3122)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 3122)
  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-031   mEosFP-M159A(G-H)//Esp3I
FEATURES             Location/Qualifiers
     source          1..3122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    740..756
                     /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(764..770,771..788,789..794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    809..830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(947..964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1118..1706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1198..1217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2015..2809
                     /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(2069..2088)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2679..2698
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2883..2901
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2939..2961)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3061..3080
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcgcag ctcagcgatc aaacccgaca tgaagatcaa cctccgaatg gaaggcaacg
       61 taaacggcca tcacttcgtg attgatggcg atggcacagg gaagccattt gaggggaaac
      121 aaagcatgga tctggaggtg aaggaaggtg gtcctctgcc ttttgccttc gacatactga
      181 ctacagcctt tcactatggg aatcgcgtgt ttgtcgagta tcccgaccat atacaggatt
      241 acttcaagca gagtttcccg aaaggctata gctgggaaag gtccctcaca tttgaggatg
      301 gagggatttg cattgcaaga aacgacatca ccatggaagg agacaccttc tacaacaagg
      361 ttcggttcca tgggacaaat ttccccgcta atggtcctgt gatgcagaag aaaaccctga
      421 agtgggaacc atccactgag aagatgtacg tgagagatgg agtactgacg ggtgacattc
      481 acgctgcact gctgttggag ggcaatgccc actatcggtg tgattttcgc actacctaca
      541 aggccaaaga gaaaggcgtc aaactccctg gataccacct tgtcgatcat tgcatcgaga
      601 tcctttctca cgacaaggac tacaacaagg tgaagctgta tgaacatgcc gttgctcact
      661 ctggattgcc agacaatgca aggcgttcag gccgagacga cctgctagcg ctcgcttgga
      721 tccgaattca aacctgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
      781 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
      841 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
      901 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
      961 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1021 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1081 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1141 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1201 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1261 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1321 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1381 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1441 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1501 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1561 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1621 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1681 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1741 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1801 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1861 aaacttggtc tgacagccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc
     1921 ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag
     1981 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     2041 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     2101 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     2161 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     2221 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     2281 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatccc accttgctcc
     2341 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     2401 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     2461 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     2521 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     2581 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     2641 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     2701 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     2761 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat ccgcgcacat
     2821 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     2881 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     2941 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3001 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     3061 cggcatcaga gcagattgta ctgagagaaa cccaattggg taccgagctc cctgcaggaa
     3121 gc
//
