LOCUS       sequence-71728-e        3695 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Supplies the selection marker (neomycin resistance) and IRES with 
            flanked by AttR2 and AttL3 sites for Gateway cloning recombination 
            reaction. (This corresponds to a module 3 plasmid)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3695)
  AUTHORS   Geiling B, Vandal G, Posner AR, de Bruyns A, Dutchak KL, Garnett S, 
            Dankort D
  TITLE     A modular lentiviral and retroviral construction system to rapidly 
            generate vectors for gene expression and gene knockdown in vitro and
            in vivo.
  JOURNAL   PLoS One. 2013 Oct 11;8(10):e76279. doi: 
            10.1371/journal.pone.0076279.
  PUBMED    24146852
REFERENCE   2  (bases 1 to 3695)
  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"; journalName: "PLoS One.";
            date: "2013-10-11"; volume: "8(10):e76279. doi"; pages: " 
            10.1371/journal.pone.0076279"
COMMENT     Sequence Label: pBEG R2-iNeo-L3   #BGV207
FEATURES             Location/Qualifiers
     source          1..3695
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    178..194
                     /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)."
     primer_bind     183..205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     202..218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     202..218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(249..373)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    386..958
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(553..570)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     780..799
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             971..1765
                     /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(1025..1044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1635..1654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(1802..1897)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1926..1945)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1927..1945)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1952..1969)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1952..1968)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1961..1983)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2175..2993)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSSRPRLNSNMDADLYGYKWARDNVGQSGATIY*LY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPG
                     KTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVG
                     IADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2898..2917
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2985..3004
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3110..3654
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3555..3574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccgcggc tcgtatgttg tgtggaattg
      181 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc actagcagat
      241 ctgccggcca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
      301 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct
      361 tgtacaaagt ggttcgataa ttccgcccct ctccctcccc cccccctaac gttactggcc
      421 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
      481 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
      541 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
      601 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
      661 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
      721 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
      781 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
      841 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
      901 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc
      961 ttgatccacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     1021 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     1081 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     1141 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     1201 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     1261 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     1321 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     1381 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     1441 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     1501 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     1561 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     1621 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     1681 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     1741 tcgccttctt gacgagttct tctgaggatc acgcgtctcg agatatcggc gcgcccccgg
     1801 gcaactttat tatacaaagt tggcattata aaaaagcatt gctcatcaat ttgttgcaac
     1861 gaacaggtca ctatcagtca aaataaaatc attatttact agtgttaact tcgatcccca
     1921 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     1981 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2041 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2101 gcgaatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa
     2161 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     2221 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     2281 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     2341 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     2401 ccatgagtga cgactgaatc cggtgagaat ggcaaaaggt tatgcatttc tttccagact
     2461 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     2521 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     2581 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     2641 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccagggat cgcagtggtg
     2701 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     2761 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     2821 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcaata gattgtcgca
     2881 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     2941 gaatttaatc gcggcctcga cgagcaagac gtttcccgtt gaatatggct cataacaccc
     3001 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     3061 tgtgcaatgt aacatcagag attttgagac actcgacaag atgatcttct tgagatcgtt
     3121 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
     3181 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt
     3241 caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct
     3301 ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac
     3361 tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata
     3421 cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa
     3481 acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac
     3541 gagggagccg ccaggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac
     3601 tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc
     3661 tttgccgcgg ccctctcact tccctcgcga aagct
//
