LOCUS       sequence-249618-        2501 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 2501)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 2501)
  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-038   Lyn(D-G)//Esp3I
FEATURES             Location/Qualifiers
     source          1..2501
                     /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    119..135
                     /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(143..149,150..167,168..173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    188..209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(326..343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(497..1085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(577..596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1394..2188
                     /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(1448..1467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2058..2077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2262..2280
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2318..2340)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2440..2459
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcggga cgccaccatg ggctgcatca agagcaagag gaaagacgtg gaaaacgtcg
       61 agggcggcag ccgagacgac ctgctagcgc tcgcttggat ccgaattcaa acctgaaatt
      121 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      181 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      241 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      301 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      661 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      901 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagccgga
     1261 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     1321 tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg
     1381 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     1441 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     1501 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     1561 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     1621 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     1681 agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat
     1741 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     1801 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     1861 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     1921 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     1981 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     2041 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     2101 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     2161 ctatcgcctt cttgacgagt tcttctgatc cgcgcacatt tccccgaaaa gtgccacctg
     2221 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2281 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2341 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2401 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2461 tgagagaaac ccaattgggt accgagctcc ctgcaggaag c
//
