LOCUS       sequence-249612-        2559 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 2559)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 2559)
  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-032   connector(H-K)//Esp3I
FEATURES             Location/Qualifiers
     source          1..2559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    177..193
                     /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(201..207,208..225,226..231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    246..267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(384..401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(555..1143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(635..654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1452..2246
                     /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(1506..1525)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2116..2135
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2320..2338
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2376..2398)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2498..2517
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcgagg ctctgattaa ggcgccaggt acatctccaa tacggcgtct tgcatagaga
       61 acacgattag cacaccggtt acccacgtga tgcccgaacg gacccatgtc atgccagagc
      121 gcactggtac gagacgacct gctagcgctc gcttggatcc gaattcaaac ctgaaattgt
      181 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      241 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
      301 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      361 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      421 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      481 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      541 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      601 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      661 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      721 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      781 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      841 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      901 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      961 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1021 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1081 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1141 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1201 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1261 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagccggaat
     1321 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
     1381 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag
     1441 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     1501 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     1561 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     1621 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     1681 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     1741 tgccggggca ggatctcctg tcatcccacc ttgctcctgc cgagaaagta tccatcatgg
     1801 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     1861 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1921 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1981 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     2041 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     2101 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     2161 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     2221 atcgccttct tgacgagttc ttctgatccg cgcacatttc cccgaaaagt gccacctgac
     2281 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2341 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     2401 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     2461 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     2521 agagaaaccc aattgggtac cgagctccct gcaggaagc
//
