LOCUS       sequence-71789-e        5692 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Supplies Cre recombinase and Luciferase 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 5692)
  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 5692)
  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-iCreT2ALuc-L3 #BGV208
FEATURES             Location/Qualifiers
     source          1..5692
                     /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    431..1003
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(598..615)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     825..844
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1019..2047
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIDYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             2063..2116
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2117..3769
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2183..2201)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(3810..3905)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3934..3953)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3935..3953)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3960..3977)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3960..3976)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3969..3991)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4183..5001)
                     /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     4906..4925
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4993..5012
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5118..5662
                     /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     5563..5582
                     /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 cggccgcacc ggtaggcctc gtacgcttaa ttaacggatc
      421 cggaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
      481 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
      541 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
      601 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
      661 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
      721 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
      781 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
      841 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
      901 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc
      961 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgat ctcggaccat
     1021 gtccaattta ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga
     1081 tgaggttcgc aagaacctga tggacatgtt cagggatcgc caggcgtttt ctgagcatac
     1141 ctggaaaatg cttctgtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg
     1201 gaaatggttt cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg
     1261 cggtctggca gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg
     1321 gtccgggctg ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg
     1381 aaaagaaaac gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga
     1441 tttcgaccag gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct
     1501 ggcatttctg gggattgatt ataacaccct gttacgtata gccgaaattg ccaggatcag
     1561 ggttaaagat atctcacgta ctgacggtgg gagaatgtta atccatattg gcagaacgaa
     1621 aacgctggtt agcaccgcag gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt
     1681 cgagcgatgg atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg
     1741 ggtcagaaaa aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct
     1801 ggaagggatt tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca
     1861 gagatacctg gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg
     1921 cgctggagtt tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt
     1981 catgaactat atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga
     2041 tggcgatgcc ggcggcagcg gcgagggcag aggcagcctg ctgacctgcg gcgacgtgga
     2101 ggagaaccct ggccccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta
     2161 tcctctagag gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
     2221 ggttcctgga acaattgctt ttacagatgc acatatcgag gtgaacatca cgtacgcgga
     2281 atacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
     2341 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
     2401 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
     2461 caacagtatg aacatttcgc agcctaccgt agtgtttgtt tccaaaaagg ggttgcaaaa
     2521 aattttgaac gtgcaaaaaa aattaccaat aatccagaaa attattatca tggattctaa
     2581 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
     2641 ttttaatgaa tacgattttg taccagagtc ctttgatcgt gacaaaacaa ttgcactgat
     2701 aatgaattcc tctggatcta ctgggttacc taagggtgtg gcccttccgc atagaactgc
     2761 ctgcgtcaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
     2821 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
     2881 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttttacg
     2941 atcccttcag gattacaaaa ttcaaagtgc gttgctagta ccaaccctat tttcattctt
     3001 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
     3061 gggcgcacct ctttcgaaag aagtcgggga agcggttgca aaacgcttcc atcttccagg
     3121 gatacgacaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
     3181 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
     3241 tctggatacc gggaaaacgc tgggcgttaa tcagagaggc gaattatgtg tcagaggacc
     3301 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
     3361 tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact tcttcatagt
     3421 tgaccgcttg aagtctttaa ttaaatacaa aggatatcag gtggcccccg ctgaattgga
     3481 atcgatattg ttacaacacc ccaacatctt cgacgcgggc gtggcaggtc ttcccgacga
     3541 tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga
     3601 aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg
     3661 agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat
     3721 cagagagatc ctcataaagg ccaagaaggg cggaaagtcc aaattgtaat aggatccgct
     3781 aggctcgacg atatcggcgc gcccccgggc aactttatta tacaaagttg gcattataaa
     3841 aaagcattgc tcatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     3901 tatttactag tgttaacttc gatccccaat tcgccctata gtgagtcgta ttacaattca
     3961 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4021 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4081 ccttcccaac agttgcgcag cctgaatggc gaatgagctt gcgccgtccc gtcaagtcag
     4141 cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag
     4201 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     4261 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     4321 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     4381 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     4441 caaaaggtta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     4501 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     4561 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     4621 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     4681 tgctgttttc ccagggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     4741 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     4801 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     4861 cttcccatac aatcaataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     4921 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgacg agcaagacgt
     4981 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     5041 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     5101 tcgacaagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
     5161 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
     5221 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     5281 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     5341 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5401 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     5461 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     5521 accgaaaggc aggaacagga gagcgcacga gggagccgcc aggggaaacg cctggtatct
     5581 ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc
     5641 aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc cc
//
