LOCUS       sequence-249609-        3176 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 3176)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 3176)
  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-029   mNeptune2.5(G-H)//Esp3I
FEATURES             Location/Qualifiers
     source          1..3176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    794..810
                     /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(818..824,825..842,843..848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    863..884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1001..1018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1172..1760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1252..1271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2069..2863
                     /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(2123..2142)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2733..2752
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2937..2955
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2993..3015)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3115..3134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgtctcgcag cgtcagcaaa ggcgaggagc tcatcaagga gaacatgcac accaagctgt
       61 acatggaagg tactgtgaat aaccaccact tcaaatgcac gcatgaaggt gaaggcaagc
      121 cctatgaggg cacacagacc aatcgcatca aagtggttga gggaggtcct ctgccgtttg
      181 cgttcgacat tctggccaca tgcttcatgt atggctccaa aacgttcatc aaccacactc
      241 agggcatacc cgatttcttc aagcagtctt ttcctgaagg ctttacatgg gagagagtga
      301 ccacatacga agatggaggc gtcctcactg tgacacaaga cacaagtctg caggatggat
      361 gtctcatcta caatgtgaag ctgcgtgggg tgaactttcc ctctaatggg cctgtaatgc
      421 agaagaaaac tcttgggtgg gaagcatcaa ccgagactct ttacccagct gatggagggc
      481 ttgaaggccg gtgtgacatg gctctcaagt tggtaggtgg aggacatctg cattgcaacc
      541 tgaaaaccac ctacaggagc aagaagccag caaagaacct gaagatgcca ggagtctatt
      601 ttgtggacag acgattggag cggattaagg aggccgacaa tgaaacctat gtcgagcaac
      661 acgaggttgc cgttgctagg tattgtgact tgccttccaa actggggcat aagctgaacg
      721 ggatggatga gctgtacaaa tcaggccgag acgacctgct agcgctcgct tggatccgaa
      781 ttcaaacctg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
      841 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      901 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      961 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1681 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1741 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1801 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1921 ggtctgacag ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
     1981 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
     2041 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     2101 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2161 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2221 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     2281 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2341 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tcccaccttg ctcctgccga
     2401 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2461 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     2521 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     2581 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     2641 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     2701 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     2761 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     2821 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgatccgcgc acatttcccc
     2881 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2941 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3001 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3061 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     3121 cagagcagat tgtactgaga gaaacccaat tgggtaccga gctccctgca ggaagc
//
