LOCUS       sequence-249601-        2416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA Kit # 1000000119 extension.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2416)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 2416)
  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: pRDAX-EMMI-021   IRES-D[C333G](H-I)//Esp3I
FEATURES             Location/Qualifiers
     source          1..2416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(19..593,594..596,597..605)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(186..203)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     413..432
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     terminator      693..722
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      744..1331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1232..1251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1343..1370)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1394..2209)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2117..2136
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2210..2312)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2313..2344)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aggctagata gatagccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
       61 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
      121 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
      181 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
      241 ggtagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
      301 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatagac ctgcaaaggc
      361 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
      421 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
      481 tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
      541 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
      601 caaccatggc gtcgagacga aacgaattca agcttgatat cattcaggac gagcctcaga
      661 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
      721 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      841 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1201 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1261 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     1321 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     1381 atttgggccc tcattagaaa aactcatcga gcatcaagtg aaactgcaat ttattcatat
     1441 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1501 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     1561 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     1621 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     1681 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcaccaacc aaaccgttat
     1741 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gccgttaaaa ggacaattac
     1801 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     1861 ctgaatcagg atattcttct aatacctgga atgctgtttt ccctgggatc gcagtggtga
     1921 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     1981 ccgtcagcca gtttagcctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     2041 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     2101 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     2161 aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcatt ttagcttcct
     2221 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     2281 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt
     2341 gactttctgc tatggaggtc aggtatgatt taaatggtca gtattgagcg atatctagag
     2401 aattcgtcac gtctcg
//
