LOCUS       sequence-274688-        3974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides IRES-BSD-pA cassette on Gateway entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3974)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3974)
  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-IRES-BSD-pA
FEATURES             Location/Qualifiers
     source          1..3974
                     /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..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(765..1339,1340..1342,1343..1351)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(932..949)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1159..1178
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     polyA_signal    1864..1998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1901..1920)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1955..1974
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2083..2178)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2215..2234)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2216..2234)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2239..2255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2239..2255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2357..2376)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2368..3177
                     /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(2435..2454)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3324..3912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3813..3832
                     /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 aaagtgggtt taaaccgccc cccccccctc tccctccccc
      781 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
      841 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
      901 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
      961 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     1021 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     1081 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     1141 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     1201 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     1261 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     1321 tgaaaaacac gatgataata tggccacaac catgcctttg tctcaagaag aatccaccct
     1381 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     1441 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     1501 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     1561 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     1621 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     1681 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     1741 ttaactagat aagtaatgat cataatcagc catatcacat ctgtagaggt tttacttgct
     1801 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1861 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1921 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1981 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2041 tgcgtattgg gcgctcttcc gcttcctcgc tcacggcgcg cccaacttta ttatacaaag
     2101 ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     2161 aaaataaaat cattatttgg agctccatgg tagcgttaac gcggccgcga tatcccctat
     2221 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2281 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     2341 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2401 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2461 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2521 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2581 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2641 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     2701 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2761 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2821 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2881 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2941 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     3001 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     3061 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     3121 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     3181 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     3241 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3301 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3361 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3421 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3481 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3541 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3601 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3661 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3721 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3781 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3841 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3901 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3961 tttgctcaca tgtt
//
