LOCUS       sequence-71723-e        3324 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Supplies the selection marker (blasticidin 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 3324)
  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 3324)
  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-iBlast-L3   #BGV204
FEATURES             Location/Qualifiers
     source          1..3324
                     /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..959
                     /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"
     protein_bind    complement(1431..1526)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1555..1574)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1556..1574)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1581..1598)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1581..1597)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1590..1612)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1804..2622)
                     /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     2527..2546
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2614..2633
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2739..3283
                     /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     3184..3203
                     /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 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag
      961 cttgatccac catgaaaaca tttaacattt ctcaacagga tctagaatta gtagaagtag
     1021 cgacagagaa gattacaatg ctttatgagg ataataaaca tcatgtggga gcggcaattc
     1081 gtacgaaaac aggagaaatc atttcggcag tacatattga agcgtatata ggacgagtaa
     1141 ctgtttgtgc agaagccatt gcgattggta gtgcagtttc gaatggacaa aaggattttg
     1201 acacgattgt agctgttaga cacccttatt ctgacgaagt agatagaagt attcgagtgg
     1261 taagtccttg tggtatgtgt agggagttga tttcagacta tgcaccagat tgttttgtgt
     1321 taatagaaat gaatggcaag ttagtcaaaa ctacgattga agaactcatt ccactcaaat
     1381 atacccgaaa ttaaggatca cgcgtctcga gatatcggcg cgcccccggg caactttatt
     1441 atacaaagtt ggcattataa aaaagcattg ctcatcaatt tgttgcaacg aacaggtcac
     1501 tatcagtcaa aataaaatca ttatttacta gtgttaactt cgatccccaa ttcgccctat
     1561 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1621 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1681 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgagct
     1741 tgcgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
     1801 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     1861 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     1921 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     1981 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     2041 gactgaatcc ggtgagaatg gcaaaaggtt atgcatttct ttccagactt gttcaacagg
     2101 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     2161 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     2221 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     2281 atattcttct aatacctgga atgctgtttt cccagggatc gcagtggtga gtaaccatgc
     2341 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     2401 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     2461 aaacaactct ggcgcatcgg gcttcccata caatcaatag attgtcgcac ctgattgccc
     2521 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     2581 cggcctcgac gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     2641 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     2701 acatcagaga ttttgagaca ctcgacaaga tgatcttctt gagatcgttt tggtctgcgc
     2761 gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc
     2821 tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     2881 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     2941 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     3001 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     3061 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     3121 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     3181 caggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     3241 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     3301 cctctcactt ccctcgcgaa agct
//
