LOCUS       40924_16910        6690 bp DNA     circular SYN 01-JAN-1980
DEFINITION  IRES-containing vector for expressing two genes in mammalian cells 
            from the same bicistronic transcript.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6690)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6690)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        154..1335
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     intron          1475..1607
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1652..1670
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1670..1692
                     /label=MCS A
                     /note="MCS A"
                     /note="multiple cloning site"
     misc_feature    1737..2323
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2332..2355
                     /label=MCS B
                     /note="MCS B"
                     /note="multiple cloning site"
     polyA_signal    complement(2394..2515)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2701..3156
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3273..3630
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3681..4472
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4539..4587
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        4893..4997
                     /label=AmpR promoter
     CDS             4998..5855
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6029..6617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagtaattca tacaaaagga ctcgcccctg ccttggggaa tcccagggac cgtcgttaaa
       61 ctcccactaa cgtagaaccc agagatcgct gcgttcccgc cccctcaccc gcccgctctc
      121 gtcatcactg aggtggagaa gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc
      181 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
      241 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
      301 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
      361 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
      421 ttacgggtta tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg
      481 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
      541 ggagcccctt cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg ccgccgcgtg
      601 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
      661 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
      721 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
      781 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
      841 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
      901 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
      961 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     1021 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     1081 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     1141 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     1201 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     1261 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     1321 tttcaggtgt cgtgaactag aagctttatt gcggtagttt atcacagtta aattgctaac
     1381 gcagtcagtg cttctgacac aacagtctcg aacttaagct gcagtgactc tcttaaggta
     1441 gccttgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     1501 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1561 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     1621 gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg ctagcctcga
     1681 gaattcacgc gtcgagcatg catctagggc ggccaattcc gcccctctcc cccccccccc
     1741 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     1801 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     1861 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     1921 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     1981 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     2041 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2101 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2161 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     2221 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2281 gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccgggatc ctctagagtc
     2341 gacccgggcg gccgcttccc tttagtgagg gttaatgctt cgagcagaca tgataagata
     2401 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2461 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2521 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     2581 caagtaaaac ctctacaaat gtggtaaaat ccgataagga tcgatccggg ctggcgtaat
     2641 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2701 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2761 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2821 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2881 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2941 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3001 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3061 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3121 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     3181 gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata
     3241 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     3301 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3361 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3421 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3481 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3541 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3601 ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg
     3661 aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3721 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3781 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3841 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3901 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3961 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4021 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4081 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4141 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4201 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4261 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4321 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4381 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4441 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4501 cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat
     4561 tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca
     4621 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4681 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4741 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4801 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4861 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4921 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4981 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5041 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5101 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5161 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5221 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5281 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5341 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5401 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5461 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5521 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5581 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5641 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5701 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5761 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5821 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5881 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5941 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6001 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6061 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6661 ggccttttgc tcacatggct cgacagatct
//
