LOCUS       pTandem-1.        3813 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian vector for the co-expression of two genes from a 
            bicistronic mRNA using the CMV promoter and an IRES.
ACCESSION   .
VERSION     .
KEYWORDS    pTandem-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3813)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3813)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          679..842
     promoter        911..929
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             961..983
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             993..995
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             999..1016
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             1026..1070
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             1086..1103
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             1122..1136
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             1233..1265
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             1272..1295
                     /label=8xHis
                     /note="8xHis affinity tag"
     misc_feature    1321..1805
                     /label=IRES
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1806..1848
                     /label=MCS 2
                     /note="MCS 2"
                     /note="multiple cloning site 2"
     polyA_signal    1913..1968
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      2056..2103
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(2275..2862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2878..3735)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3736..3806)
                     /label=AmpR promoter
     promoter        3812..3813
                     /label=CMV
                     /note="Human cytomegalovirus immediate early
                     enhancer/promoter"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
      361 tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga cgtcgtttag tgaaccgtca gatcactaga tgctttattg
      601 cggtagttta tcacagttaa attgctaacg ccagtctcga acttaacgtg cagaagttgg
      661 tcgtgaggca ctgggcaggt aagtatcggg ccctttgtgc ggggggagcg gctcggggct
      721 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      781 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      841 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc
      901 ggaccgaaat taatacgact cactataggg agtccacaac ggtttccctc tagaaataat
      961 tttgtttaac tttaagaagg agatatgcca ccatggcaca ccatcaccac catcactctt
     1021 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg ccaccgcctt
     1081 ctggtctggt cccccggggc agcgcaggtt ctggtacgat tgatgacgac gacaagagtc
     1141 aattgatatc ggcgcgccat gcattaatta atttaaatgg cccgtacggc cggtcaccgg
     1201 tagcggccgc acagctgtat acacgtgcaa gccagccaga actcgccccg gaagaccccg
     1261 aggatctcga gcaccaccat caccatcacc atcactaagt gattaacctc aggactagtg
     1321 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1381 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     1441 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     1501 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     1561 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     1621 agttgtggaa agagtcaaat ggctcacctc aagcgtattc aacaaggggc tgaaggatgc
     1681 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     1741 tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     1801 tgaaagttta aacgctagcc aagcgcttgg ttaacgctga gcatcgatct ttttccctct
     1861 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     1921 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     1981 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     2041 tatgcccata tgtaactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2101 ttgctgaaac atgcgaacgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2281 taggctccgc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2341 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2401 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2461 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2521 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2581 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2641 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2701 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2761 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2821 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgttacca
     2881 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2941 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3001 tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3061 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3121 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3181 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3241 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3301 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3361 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3421 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3481 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3541 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3601 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3661 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3721 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3781 tctcctgagc ggatacatat ttgaaggatc cct
//
