LOCUS       pRetroX-PTuner_I        6283 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Retroviral vector for fusing a destabilization domain to the 
            N-terminus of a partner protein, with co-expression of ZsGreen1.
ACCESSION   .
VERSION     .
KEYWORDS    pRetroX-PTuner IRES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6283)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6283)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    655..1012
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     intron          1075..1278
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of
                     human EF-1-alpha"
     exon            1279..1308
                     /label=EF-1-alpha exon
                     /note="truncated exon 2 from human EF-1-alpha, beginning 
                     with a GT-to-CC mutation that reduces splicing efficiency"
     CDS             1368..1691
                     /label=DD
                     /note="destabilization domain that can be stabilized by
                     Shield1 in the ProteoTuner(TM) system"
     misc_feature    1694..1719
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    1723..2302
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2297..2989
                     /label=ZsGreen1
                     /note="Zoanthus green fluorescent protein"
     LTR             3046..3639
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     primer_bind     complement(3655..3671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3679..3695)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3703..3733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3748..3769)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4057..4645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4819..5676)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5677..5781)
                     /label=AmpR promoter
     primer_bind     6255..6271
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag
      121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      241 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      301 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      361 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgggctcaat
      421 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg
      481 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggcct cgctgttcct
      541 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      601 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
      661 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      721 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      781 gttcggaaca cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg
      841 tggcccgacc tgagtccaaa aatcccgatc gttttggact ctttggtgca ccccccttag
      901 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
      961 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg
     1021 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaatacg cgtgtcgagc
     1081 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     1141 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     1201 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     1261 ttttttcttc catttcagcc gtcgtgaaaa ctacccctaa aagccaaaaa aatcgaattc
     1321 ccaaacttaa gcttggtacc gagctcggat ctcacgtggg cccgcggatg ggagtgcagg
     1381 tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgtgtgg
     1441 tgcactacac cgggatgctt gaagatggaa agaaagtcga ttcctcccgg gacagaaaca
     1501 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg
     1561 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg
     1621 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc
     1681 taaaaccgga accgcggccg cagatctatc gatggatccg tgcccctctc cctccccccc
     1741 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     1801 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     1861 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     1921 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     1981 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     2041 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     2101 tgtggaaaga gtcaaatggc tcacctcaag cgtattcaac aaggggctga aggatgccca
     2161 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     2221 ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     2281 aaaacacgat gataatatgg cccagtccaa gcacggcctg accaaggaga tgaccatgaa
     2341 gtaccgcatg gagggctgcg tggacggcca caagttcgtg atcaccggcg agggcatcgg
     2401 ctaccccttc aagggcaagc aggccatcaa cctgtgcgtg gtggagggcg gccccttgcc
     2461 cttcgccgag gacatcttgt ccgccgcctt catgtacggc aaccgcgtgt tcaccgagta
     2521 cccccaggac atcgtcgact acttcaagaa ctcctgcccc gccggctaca cctgggaccg
     2581 ctccttcctg ttcgaggacg gcgccgtgtg catctgcaac gccgacatca ccgtgagcgt
     2641 ggaggagaac tgcatgtacc acgagtccaa gttctacggc gtgaacttcc ccgccgacgg
     2701 ccccgtgatg aagaagatga ccgacaactg ggagccctcc tgcgagaaga tcatccccgt
     2761 gcccaagcag ggcatcttga agggcgacgt gagcatgtac ctgctgctga aggacggtgg
     2821 ccgcttgcgc tgccagttcg acaccgtgta caaggccaag tccgtgcccc gcaagatgcc
     2881 cgactggcac ttcatccagc acaagctgac ccgcgaggac cgcagcgacg ccaagaacca
     2941 gaagtggcac ctgaccgagc acgccatcgc ctccggctcc gccttgccct gagcggcttc
     3001 gaggataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     3061 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     3121 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     3181 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     3241 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3301 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     3361 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3421 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3481 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     3541 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3601 tctgagtgat tgactacccg tcagcggggg tctttcattg aattcgtaat catggtcata
     3661 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3721 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3781 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3841 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3901 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3961 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4021 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4081 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4141 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4201 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4261 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4321 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4381 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4441 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4501 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4561 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4621 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4681 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4741 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4801 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4861 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4921 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4981 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5041 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5101 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5161 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5221 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5281 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5341 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5401 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5461 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5521 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5581 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5641 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5701 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5761 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5821 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5881 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5941 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6001 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6061 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6121 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6181 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6241 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt atg
//
