LOCUS       sequence-71686-e        3779 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Contains the Gateway selection cassette (ccdB ChlorR) flanked by 
            AttR1 and AttR3 sites for the creation of Destination vectors .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3779)
  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 3779)
  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 R1-ChlorR-R3   #BGV148
FEATURES             Location/Qualifiers
     source          1..3779
                     /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    249..373
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(404..409,410..427,428..434)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             488..1147
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(554..573)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1489..1794
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1673..1692
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1859..1982)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2021..2040)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2022..2040)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2047..2064)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2047..2063)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2056..2078)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2270..3088)
                     /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     2993..3012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3080..3099
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3205..3749
                     /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     3650..3669
                     /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 ctgctagcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
      301 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
      361 tccagtcact atggccggcg cggccgcatt aggcacccca ggctttacac tttatgcttc
      421 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
      481 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
      541 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
      601 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
      661 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
      721 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
      781 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
      841 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
      901 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
      961 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     1021 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     1081 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1141 ggcgtaatct agaggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     1201 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     1261 tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     1321 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     1381 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     1441 ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag
     1501 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     1561 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     1621 aaagtccccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     1681 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     1741 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     1801 ggctccctta tacacagcca gtctgcaggt cgacgatatc ggcgcgccaa ccccggccca
     1861 tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta
     1921 atttaatata ttgatattta tatcatttta cgtttctcgt tcaactttat tatacatagt
     1981 tggctagtgg cgttaacggt gaacttcgat ccccaattcg ccctatagtg agtcgtatta
     2041 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2101 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2161 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgagcttgcg ccgtcccgtc
     2221 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact
     2281 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     2341 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2401 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2461 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2521 agaatggcaa aaggttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2581 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2641 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2701 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2761 cctggaatgc tgttttccca gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2821 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2881 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2941 catcgggctt cccatacaat caatagattg tcgcacctga ttgcccgaca ttatcgcgag
     3001 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgagc
     3061 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     3121 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     3181 gagacactcg acaagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc
     3241 tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc
     3301 tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta
     3361 gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg
     3421 ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg
     3481 cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct
     3541 acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca
     3601 ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg ggaaacgcct
     3661 ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat
     3721 gcttgtcagg ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccc
//
