LOCUS       sequence-249613-        2568 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 2568)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 2568)
  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-033   connector(M-W)//Esp3I
FEATURES             Location/Qualifiers
     source          1..2568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    186..202
                     /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(210..216,217..234,235..240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    255..276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(393..410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(564..1152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(644..663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1461..2255
                     /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(1515..1534)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2125..2144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2329..2347
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2385..2407)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2507..2526
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcatca cccgattctc atcatggaga cacatcagga catctccaat accgccatgg
       61 aaacgcacca agatgcgact gctcttctgc cagtgacggc attcgtctgc aagattaacg
      121 gcatacagaa agaaacgatg agacgacctg ctagcgctcg cttggatccg aattcaaacc
      181 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      241 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      361 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      421 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      481 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      541 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      601 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      661 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      721 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      781 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      841 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      901 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      961 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1021 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1081 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1141 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1201 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1261 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1321 agccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     1381 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     1441 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     1501 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     1561 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     1621 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     1681 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     1741 tgggcgaagt gccggggcag gatctcctgt catcccacct tgctcctgcc gagaaagtat
     1801 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     1861 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     1921 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     1981 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     2041 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     2101 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     2161 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     2221 tcgccttcta tcgccttctt gacgagttct tctgatccgc gcacatttcc ccgaaaagtg
     2281 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2341 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2401 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2461 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     2521 attgtactga gagaaaccca attgggtacc gagctccctg caggaagc
//
