LOCUS       sequence-249619-        2516 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 2516)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 2516)
  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-039   LifeAct(D-G)//Esp3I
FEATURES             Location/Qualifiers
     source          1..2516
                     /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"
     CDS             18..68
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     protein_bind    134..150
                     /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(158..164,165..182,183..188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    203..224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(341..358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(512..1100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(592..611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1409..2203
                     /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(1463..1482)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2073..2092
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2277..2295
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2333..2355)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2455..2474
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcggga cgccaccatg ggcgtggccg acctgatcaa gaagttcgag agcatcagca
       61 aggaggaggg cgggtcaggc ggcagccgag acgacctgct agcgctcgct tggatccgaa
      121 ttcaaacctg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
      181 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      241 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      301 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      361 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      421 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1021 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1081 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1141 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1261 ggtctgacag ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
     1321 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
     1381 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     1441 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     1501 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     1561 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     1621 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     1681 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tcccaccttg ctcctgccga
     1741 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     1801 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     1861 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     1921 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     1981 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     2041 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     2101 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     2161 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgatccgcgc acatttcccc
     2221 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2281 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2341 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2401 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2461 cagagcagat tgtactgaga gaaacccaat tgggtaccga gctccctgca ggaagc
//
