LOCUS       sequence-71726-e        3927 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Supplies the selection marker (hygromycin 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 3927)
  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 3927)
  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-iHygro-L3   #BGV206
FEATURES             Location/Qualifiers
     source          1..3927
                     /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    390..962
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(557..574)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     784..803
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             967..1992
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             967..1992
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(2034..2129)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2158..2177)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2159..2177)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2184..2201)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2184..2200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2193..2215)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2407..3225)
                     /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     3130..3149
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3217..3236
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3342..3886
                     /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     3787..3806
                     /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 ggtgctagcg cggccgcccc ccctctccct cccccccccc taacgttact
      421 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
      481 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
      541 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
      601 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
      661 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
      721 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
      781 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
      841 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
      901 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
      961 aagcttatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     1021 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     1081 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     1141 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     1201 gacattgggg aatttagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     1261 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     1321 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     1381 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     1441 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     1501 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     1561 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     1621 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     1681 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     1741 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     1801 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     1861 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     1921 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     1981 gcaaaggaat gaggatctcc taggctcgag ttaagggtgg gcgcgccccc gggcaacttt
     2041 attatacaaa gttggcatta taaaaaagca ttgctcatca atttgttgca acgaacaggt
     2101 cactatcagt caaaataaaa tcattattta ctagtgttaa cttcgatccc caattcgccc
     2161 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2221 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2281 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatga
     2341 gcttgcgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa
     2401 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     2461 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     2521 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     2581 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     2641 gacgactgaa tccggtgaga atggcaaaag gttatgcatt tctttccaga cttgttcaac
     2701 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     2761 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     2821 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     2881 aggatattct tctaatacct ggaatgctgt tttcccaggg atcgcagtgg tgagtaacca
     2941 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     3001 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     3061 cagaaacaac tctggcgcat cgggcttccc atacaatcaa tagattgtcg cacctgattg
     3121 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     3181 tcgcggcctc gacgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     3241 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     3301 gtaacatcag agattttgag acactcgaca agatgatctt cttgagatcg ttttggtctg
     3361 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     3421 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     3481 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     3541 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     3601 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     3661 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     3721 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     3781 cgccagggga aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga
     3841 gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc
     3901 ggccctctca cttccctcgc gaaagct
//
