LOCUS       sequence-274701-        5171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides mouse ER protein tag N-terminal to IRES-puro-pA cassette on
            Gateway entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5171)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 5171)
  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"
COMMENT     Sequence Label: pENTR-R2L3-cmER-IRESpuro-pA
FEATURES             Location/Qualifiers
     source          1..5171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..748)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(1762..2336,2337..2339,2340..2348)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1929..1946)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2156..2175
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2349..2948
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMVGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2349..2368)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2845..2865
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3029..3046)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3035..3259
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(3280..3375)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3412..3431)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3413..3431)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3436..3452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3436..3452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3554..3573)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3565..4374
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3632..3651)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4521..5109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5010..5029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggtca tgagcactag tcgaaatgaa atgggtgctt
      781 caggagacat gagggctgcc aacctttggc caagccctct tgtgattaag cacactaaga
      841 agaatagccc tgccttgtcc ttgacagctg accagatggt cagtgccttg ttggatgctg
      901 aaccgcccat gatctattct gaatatgatc cttctagacc cttcagtgaa gcctcaatga
      961 tgggcttatt gaccaaccta gcagataggg agctggttca tatgatcaac tgggcaaaga
     1021 gagtgccagg ctttggggac ttgaatctcc atgatcaggt ccaccttctc gagtgtgcct
     1081 ggctggagat tctgatgatt ggtctcgtct ggcgctccat ggaacacccg gggaagctcc
     1141 tgtttgctcc taacttgctc ctggacagga atcaaggtaa atgtgtggaa ggcatggtgg
     1201 agatctttga catgttgctt gctacgtcaa gtcggttccg catgatgaac ctgcagggtg
     1261 aagagtttgt gtgcctcaaa tccatcattt tgcttaattc cggagtgtac acgtttctgt
     1321 ccagcacctt gaagtctctg gaagagaagg accacatcca ccgtgtcctg gacaagatca
     1381 cagacacttt gatccacctg atggccaaag ctggcctgac tctgcagcag cagcatcgcc
     1441 gcctagctca gctccttctc attctttccc atatccggca tatgagtaac aaacgcatgg
     1501 agcatctcta caacatgaaa tgcaagaacg tggtacccct ctatgacctg ctcctggaga
     1561 tgttggatgc ccaccgcctt catgccccag ccagtcgcat gggagtgccc ccagaggagc
     1621 ccagccagac ccagctggcc accaccagct ccacttcagc acattcctta caaacctact
     1681 acataccccc ggaagcagag ggcttcccca acacgatctg agcggccgct cgataagctt
     1741 gatatcgaat tccgcccccc ccccctctcc ctcccccccc cctaacgtta ctggccgaag
     1801 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     1861 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     1921 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     1981 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     2041 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     2101 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     2161 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     2221 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     2281 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
     2341 ccacaaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     2401 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     2461 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     2521 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     2581 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     2641 tgagcggttc ccggctggcc gcgcagcaac agatggtagg cctcctggcg ccgcaccggc
     2701 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     2761 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     2821 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     2881 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     2941 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     3001 catcgatgat ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3061 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3121 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3181 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3241 gatgcggtgg gctctatggc ttctgaggcg gaaagaaccc aactttatta tacaaagttg
     3301 gcattataaa aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     3361 ataaaatcat tatttggagc tccatggtag cgttaacgcg gccgcgatat cccctatagt
     3421 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     3481 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     3541 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     3601 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     3661 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     3721 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     3781 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     3841 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     3901 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     3961 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     4021 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     4081 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     4141 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     4201 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     4261 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     4321 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     4381 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     4441 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     4501 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4561 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4621 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4681 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4741 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4801 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4861 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4921 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4981 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5041 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5101 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5161 gctcacatgt t
//
