LOCUS       sequence-71730-e        3480 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Supplies the selection marker (puromycin 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 3480)
  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 3480)
  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-iPuro-L3   #BGV205
FEATURES             Location/Qualifiers
     source          1..3480
                     /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             963..1562
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(963..982)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1459..1479
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(1599..1694)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1723..1742)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1724..1742)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1749..1766)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1749..1765)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1758..1780)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1972..2790)
                     /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     2695..2714
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2782..2801
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2907..3451
                     /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     3352..3371
                     /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 ttatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     1021 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     1081 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     1141 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     1201 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     1261 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     1321 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     1381 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     1441 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     1501 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     1561 gaggatcacg cgtctcgaga tatcggcgcg cccccgggca actttattat acaaagttgg
     1621 cattataaaa aagcattgct catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     1681 taaaatcatt atttactagt gttaacttcg atccccaatt cgccctatag tgagtcgtat
     1741 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1801 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     1861 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgagcttg cgccgtcccg
     1921 tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg attagaaaaa
     1981 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     2041 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     2101 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     2161 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     2221 tgagaatggc aaaaggttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     2281 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     2341 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     2401 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     2461 tacctggaat gctgttttcc cagggatcgc agtggtgagt aaccatgcat catcaggagt
     2521 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     2581 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     2641 cgcatcgggc ttcccataca atcaatagat tgtcgcacct gattgcccga cattatcgcg
     2701 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacga
     2761 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     2821 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     2881 ttgagacact cgacaagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc
     2941 tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac
     3001 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     3061 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     3121 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     3181 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     3241 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     3301 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca ggggaaacgc
     3361 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     3421 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
//
