LOCUS       sequence-71697-e        3628 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Supplies the selection marker (dsRed) 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 3628)
  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 3628)
  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-dsRed-L3   #BGV203
FEATURES             Location/Qualifiers
     source          1..3628
                     /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    join(425..999,1000..1002,1003..1011)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(592..609)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     819..838
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1027..1704
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MTSSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(1207..1227)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1619..1642
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(1746..1841)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1870..1889)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1871..1889)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1896..1913)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1896..1912)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1905..1927)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2119..2937)
                     /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     2842..2861
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2929..2948
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3054..3598
                     /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     3499..3518
                     /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 cttcaccttg gatccgccgg cgaattccgc
      421 cccccccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
      481 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
      541 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
      601 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
      661 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
      721 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
      781 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
      841 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
      901 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
      961 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catggccaga
     1021 tccgtcatga cctcctccga gaacgtcatc accgagttca tgcgcttcaa ggtgcgcatg
     1081 gagggcaccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1141 gagggccaca acaccgtgaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     1201 gacatcctgt ccccccagtt ccagtacggc tccaaggtgt acgtgaagca ccccgccgac
     1261 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1321 ttcgaggacg gcggcgtggc gaccgtgacc caggactcct ccctgcagga cggctgcttc
     1381 atctacaagg tgaagttcat cggcgtgaac ttcccctccg acggccccgt gatgcagaag
     1441 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1501 ggcgagaccc acaaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1561 tccatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacgccaag
     1621 ctggacatca cctcccacaa cgaggactac accatcgtgg agcagtacga gcgcaccgag
     1681 ggccgccacc acctgttcct gtagagatct cctaggctcg agttaagggt gggcgcgccc
     1741 ccgggcaact ttattataca aagttggcat tataaaaaag cattgctcat caatttgttg
     1801 caacgaacag gtcactatca gtcaaaataa aatcattatt tactagtgtt aacttcgatc
     1861 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     1921 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     1981 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2041 aatggcgaat gagcttgcgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa
     2101 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2161 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2221 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     2281 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2341 atcaccatga gtgacgactg aatccggtga gaatggcaaa aggttatgca tttctttcca
     2401 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2461 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     2521 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     2581 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccag ggatcgcagt
     2641 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     2701 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     2761 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc aatagattgt
     2821 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     2881 gttggaattt aatcgcggcc tcgacgagca agacgtttcc cgttgaatat ggctcataac
     2941 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     3001 atcttgtgca atgtaacatc agagattttg agacactcga caagatgatc ttcttgagat
     3061 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     3121 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     3181 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     3241 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     3301 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     3361 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     3421 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     3481 gcacgaggga gccgccaggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3541 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     3601 cggctttgcc gcggccctct cacttccc
//
