LOCUS       pCI-TPI-PTC48-HBB      8170 bp    DNA     circular SYN 23-JUN-2000
DEFINITION  Cloning vector pCI-neo, mammalian expression vector, complete
            sequence.
ACCESSION   U47120
VERSION     U47120.2  GI:8677398
KEYWORDS    .
SOURCE      Cloning vector pCI-neo (unknown)
  ORGANISM  Cloning vector pCI-neo
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 5472)
  AUTHORS   Brondyk,W.H.
  TITLE     New Mammalian Expression Vector with a selectable marker: pCI-neo
  JOURNAL   Promega Notes 51, 10-14 (1995)
REFERENCE   2  (bases 1 to 5472)
  AUTHORS   Brondyk,W.H. and Groskreutz,D.J.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JAN-1996) Melinda Brady, R&D, Promega Corporation,
            5445 East Cheryl Parkway, Madison, WI 53711, USA
REFERENCE   3  (bases 1 to 5472)
  AUTHORS   Kamal,S.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JUN-2000) Technical Writing, Promega Corporation,
            5445 East Cheryl Parkway, Madison, WI 53711, USA
   REMARK   Sequence updated by submitter
COMMENT     On Jun 23, 2000 this sequence version replaced gi:1197674.
FEATURES             Location/Qualifiers
     regulatory      1..659
                     /regulatory_class="enhancer"
                     /note="Cytomegalovirus immediate-early enhancer"
     Vector          join(3830..8170,1..1085)
                     /origin="CVU47120 (5.472bp circular)Target vector: from
                     NotI cut at 1131^1132 to NheI cut at 1085^1086,
                     5.426bpOrientation: originalPlus strand 5':   no
                     changePlus strand 3':   no changeMinus strand 5':  no
                     changeMinus strand 3':  no change"
     regulatory      669..750
                     /regulatory_class="promoter"
                     /note="Cytomegalovirus immediate-early promoter"
     Intron          890..1022
                     /note="chimeric intron"
     regulatory      1067..1085
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     Misc._feature   join(<3830..3835,1085..>1085)
                     /note="multiple cloning sites"
     Exon            1086..1259
                     /label=Exon1
     ORF             join(1145..1259,2192..2315,2427..2511,2586..2718,3016..3101,3377..3464,3593..3748)
                     /label=TPI
     Exon            2192..2315
                     /label=Exon2
     Mutation        2218..2220
                     /label=PTC48
     Exon            2427..2511
                     /label=Exon3
     Exon            2586..2718
                     /label=Exon4
     Exon            3016..3101
                     /label=Exon5
     Exon            3377..3464
                     /label=Exon6
     Exon            3593..3678
                     /label=Exon7
     Binding_site    3679..3827
                     /label=HBB_long
     regulatory      3838..3856
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     regulatory      3865..4086
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyadenylation signal"
     Replication_ori 4181..4636
                     /note="phage f1 region"
     regulatory      4700..5116
                     /regulatory_class="promoter"
                     /note="SV40 enhancer and early promoter - neomycin
                     selectable marker"
     Replication_ori 5014..5079
                     /note="SV40 minimum origin of replication - neomycin
                     selectable marker"
     Gene            5161..5955
                     /gene="NeoR"
     CDS             5161..5955
                     /gene="NeoR"
                     /function="neomycin resistance"
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /protein_id="AAA88781.1"
                     /db_xref="GI:1197675"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      6019..6067
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyadenylation signal - neomycin
                     selectable marker"
     Gene            6478..7338
                     /gene="ampR"
     CDS             6478..7338
                     /gene="ampR"
                     /function="ampicillin resistance"
                     /codon_start=1
                     /product="beta-lactamase"
                     /protein_id="AAA88782.1"
                     /db_xref="GI:1197676"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
ORIGIN
        1 TCAATATTGG CCATTAGCCA TATTATTCAT TGGTTATATA GCATAAATCA ATATTGGCTA
       61 TTGGCCATTG CATACGTTGT ATCTATATCA TAATATGTAC ATTTATATTG GCTCATGTCC
      121 AATATGACCG CCATGTTGGC ATTGATTATT GACTAGTTAT TAATAGTAAT CAATTACGGG
      181 GTCATTAGTT CATAGCCCAT ATATGGAGTT CCGCGTTACA TAACTTACGG TAAATGGCCC
      241 GCCTGGCTGA CCGCCCAACG ACCCCCGCCC ATTGACGTCA ATAATGACGT ATGTTCCCAT
      301 AGTAACGCCA ATAGGGACTT TCCATTGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC
      361 CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTCCG CCCCCTATTG ACGTCAATGA
      421 CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC TTACGGGACT TTCCTACTTG
      481 GCAGTACATC TACGTATTAG TCATCGCTAT TACCATGGTG ATGCGGTTTT GGCAGTACAC
      541 CAATGGGCGT GGATAGCGGT TTGACTCACG GGGATTTCCA AGTCTCCACC CCATTGACGT
      601 CAATGGGAGT TTGTTTTGGC ACCAAAATCA ACGGGACTTT CCAAAATGTC GTAACAACTG
      661 CGATCGCCCG CCCCGTTGAC GCAAATGGGC GGTAGGCGTG TACGGTGGGA GGTCTATATA
      721 AGCAGAGCTC GTTTAGTGAA CCGTCAGATC ACTAGAAGCT TTATTGCGGT AGTTTATCAC
      781 AGTTAAATTG CTAACGCAGT CAGTGCTTCT GACACAACAG TCTCGAACTT AAGCTGCAGT
      841 GACTCTCTTA AGGTAGCCTT GCAGAAGTTG GTCGTGAGGC ACTGGGCAGG TAAGTATCAA
      901 GGTTACAAGA CAGGTTTAAG GAGACCAATA GAAACTGGGC TTGTCGAGAC AGAGAAGACT
      961 CTTGCGTTTC TGATAGGCAC CTATTGGTCT TACTGACATC CACTTTGCCT TTCTCTCCAC
     1021 AGGTGTCCAC TCCCAGTTCA ATTACAGCTC TTAAGGCTAG AGTACTTAAT ACGACTCACT
     1081 ATAGGCTAGC CTCGAGAATT CACGCGTGGT ACCTCTAGAC CTTCAGCGCC TCGGCTCCAG
     1141 CACCATGGcg ccctccagga agttcttcgt tgggggaaac tggaagatga acgggcggaa
     1201 gcagagtctg ggggagctca tcggcactct gaacgcggcc aaggtgccgg ccgacaccgg
     1261 taagccctcg ccgaggaggg gtctggccgg gccggggccg gggccggggc aggagtggca
     1321 gcgccctctc ccgaggcccc gaggccccga ggccggtatc cgcgcggacc tgatgcaggg
     1381 ctgtgggacg agggccgctg gggtccgggc aggggcctcg cagccgcagc cccgtcggtg
     1441 cgtcgagggg gcagggcgga gcacatgatg ccccttggac tatggggcag gtaaggacgt
     1501 tttcttccct tcggcaacct gaacgactcc cgccttccac ggaagggacc gagcccgtgc
     1561 caaacaggct gagcgatttg ggagtgagga gccatcctac cgctttcccc aacctggaaa
     1621 cagcaaagcg caaggcctct gagtcagtta ggtctctgcc acccacgggc aaaggatgct
     1681 ctcctccatc ctccttcctc cctccaccga aatcggagag ccgcgggcct gatccaaaga
     1741 ggcatcccct tctcgttcat tccccagagg cctcaataca aaccccagga gttggcccct
     1801 ctccttttgc tacaaatcct tgccttgcaa aggggaggtg aggatgggct attttagaag
     1861 ggaagcaggg ttgctccctg gagaatgctg agtctgtgag gtgcctatgc cgagaatagc
     1921 tcgaggaaat tggagcccca gctgttaaaa gagcagaggg cagggtgagg gccgtggcct
     1981 ctcaggggta tctggaaggc tcttcgagtt gagtgcagac ccagcctggg ctggaaaatg
     2041 gacaaaggtc atcttgctgg ggtgaaaagg gggagagcag aaccaagaag aagagggtga
     2101 gggctggggg gctccagggc actggttagg aattgtgggg aatgaaggct ttctttagtc
     2161 tcatccccct gtggtaccat cttgtcctca gaggtggttt gtgctccccc tactgcctaa
     2221 atcgacttcg cccggcagaa gctagatccc aagattgctg tggctgcgca gaactgctac
     2281 aaagtgacta atggggcttt tactggggag atcaggtgag atcgaggtgg agaggggtgt
     2341 gtgggaccct tccctcactt tcctcgttga ggggaaagcc acagggtggg ctccctgctg
     2401 aaccttggct tcatctcttc ctttagccct ggcatgatca aagactgcgg agccacgtgg
     2461 gtggtcctgg ggcactcaga gagaaggcat gtctttgggg agtcagatga ggttagtagc
     2521 caagagagaa gataagggat gtctttttcc aagaaggatg tctcaccaag tctgtttctc
     2581 aacagctgat tgggcagaaa gtggcccatg ctctggcaga gggactcgga gtaatcgcct
     2641 gcattgggga gaagctagat gaaagggaag ctggcatcac tgagaaggtt gttttcgagc
     2701 agacaaaggt catcgcaggt atctctggag aaagggacct ttgagcctat ccagggccac
     2761 agagactcag agggtagggt caggccctgg agcctgtctt ggtccccatg ctgatccaga
     2821 aaaggaaaaa ggggaggggg agtgacaatc tttgcttggg gcctatgact tctccagccc
     2881 caaggtagat gccacctgga aatcccccaa tgtccactag ggggcagtag gccaccgttc
     2941 ttcgtactcc ggagaacctg gctggagagc tctttcttgt tcacccttcc ctccatctgt
     3001 atctctgccc tgcagataac gtgaaggact ggagcaaggt cgtcctggcc tatgagcctg
     3061 tgtgggccat tggtactggc aagactgcaa caccccaaca ggtaaccggg cccaggagcc
     3121 ctgccctcat cccagcctgc ctcaataggt ttggacagac acagcccaca tggggcaacc
     3181 ccttatttca aagacacaga gaccttgaac ccagagacag tgacttgtcc aagggcatcc
     3241 agtccagggc ctggcttgga tcagagccct ggtactctga ctcagtcaga aaccacacta
     3301 agtgtccact ggtgccagtg atttttcctc ttagagaggc agaaaaggtc ttacttaggc
     3361 cagcttcttg ttctaggccc aggaagtaca cgagaagctc cgaggatggc tgaagtccaa
     3421 cgtctctgat gcggtggctc agagcacccg tatcatttat ggaggtgagt ggctttggtt
     3481 cccggctgag gtggagtggg ctgaggacta gactgagccc tcggacatgg aggtggggat
     3541 ggggcagact catcccattc ttgaccaagc ccttgttctg ctcccttccc aggctctgtg
     3601 actggggcaa cctgcaagga gctggccagc cagcctgatg tggatggctt ccttgtgggt
     3661 ggtgcttccc tcaagcccAT CACTTTGGCA AAGAATTCAC CCCACCAGTG CAGGCTGCCT
     3721 ATCAGAAAGT GGTGGCTGGT GTGGCTAATG CCCTGGCCCA CAAGTATCAC TAAGCTCGCT
     3781 TTCTTGCTGT CCAATTTCTA TTAAAGGTTC CTTTGTTCCC TAAGTCCGCG GCCGCTTCCC
     3841 TTTAGTGAGG GTTAATGCTT CGAGCAGACA TGATAAGATA CATTGATGAG TTTGGACAAA
     3901 CCACAACTAG AATGCAGTGA AAAAAATGCT TTATTTGTGA AATTTGTGAT GCTATTGCTT
     3961 TATTTGTAAC CATTATAAGC TGCAATAAAC AAGTTAACAA CAACAATTGC ATTCATTTTA
     4021 TGTTTCAGGT TCAGGGGGAG ATGTGGGAGG TTTTTTAAAG CAAGTAAAAC CTCTACAAAT
     4081 GTGGTAAAAT CCGATAAGGA TCGATCCGGG CTGGCGTAAT AGCGAAGAGG CCCGCACCGA
     4141 TCGCCCTTCC CAACAGTTGC GCAGCCTGAA TGGCGAATGG ACGCGCCCTG TAGCGGCGCA
     4201 TTAAGCGCGG CGGGTGTGGT GGTTACGCGC AGCGTGACCG CTACACTTGC CAGCGCCCTA
     4261 GCGCCCGCTC CTTTCGCTTT CTTCCCTTCC TTTCTCGCCA CGTTCGCCGG CTTTCCCCGT
     4321 CAAGCTCTAA ATCGGGGGCT CCCTTTAGGG TTCCGATTTA GTGCTTTACG GCACCTCGAC
     4381 CCCAAAAAAC TTGATTAGGG TGATGGTTCA CGTAGTGGGC CATCGCCCTG ATAGACGGTT
     4441 TTTCGCCCTT TGACGTTGGA GTCCACGTTC TTTAATAGTG GACTCTTGTT CCAAACTGGA
     4501 ACAACACTCA ACCCTATCTC GGTCTATTCT TTTGATTTAT AAGGGATTTT GCCGATTTCG
     4561 GCCTATTGGT TAAAAAATGA GCTGATTTAA CAAAAATTTA ACGCGAATTT TAACAAAATA
     4621 TTAACGCTTA CAATTTCCTG ATGCGGTATT TTCTCCTTAC GCATCTGTGC GGTATTTCAC
     4681 ACCGCATACG CGGATCTGCG CAGCACCATG GCCTGAAATA ACCTCTGAAA GAGGAACTTG
     4741 GTTAGGTACC TTCTGAGGCG GAAAGAACCA GCTGTGGAAT GTGTGTCAGT TAGGGTGTGG
     4801 AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG TATGCAAAGC ATGCATCTCA ATTAGTCAGC
     4861 AACCAGGTGT GGAAAGTCCC CAGGCTCCCC AGCAGGCAGA AGTATGCAAA GCATGCATCT
     4921 CAATTAGTCA GCAACCATAG TCCCGCCCCT AACTCCGCCC ATCCCGCCCC TAACTCCGCC
     4981 CAGTTCCGCC CATTCTCCGC CCCATGGCTG ACTAATTTTT TTTATTTATG CAGAGGCCGA
     5041 GGCCGCCTCG GCCTCTGAGC TATTCCAGAA GTAGTGAGGA GGCTTTTTTG GAGGCCTAGG
     5101 CTTTTGCAAA AAGCTTGATT CTTCTGACAC AACAGTCTCG AACTTAAGGC TAGAGCCACC
     5161 ATGATTGAAC AAGATGGATT GCACGCAGGT TCTCCGGCCG CTTGGGTGGA GAGGCTATTC
     5221 GGCTATGACT GGGCACAACA GACAATCGGC TGCTCTGATG CCGCCGTGTT CCGGCTGTCA
     5281 GCGCAGGGGC GCCCGGTTCT TTTTGTCAAG ACCGACCTGT CCGGTGCCCT GAATGAACTG
     5341 CAGGACGAGG CAGCGCGGCT ATCGTGGCTG GCCACGACGG GCGTTCCTTG CGCAGCTGTG
     5401 CTCGACGTTG TCACTGAAGC GGGAAGGGAC TGGCTGCTAT TGGGCGAAGT GCCGGGGCAG
     5461 GATCTCCTGT CATCTCACCT TGCTCCTGCC GAGAAAGTAT CCATCATGGC TGATGCAATG
     5521 CGGCGGCTGC ATACGCTTGA TCCGGCTACC TGCCCATTCG ACCACCAAGC GAAACATCGC
     5581 ATCGAGCGAG CACGTACTCG GATGGAAGCC GGTCTTGTCG ATCAGGATGA TCTGGACGAA
     5641 GAGCATCAGG GGCTCGCGCC AGCCGAACTG TTCGCCAGGC TCAAGGCGCG CATGCCCGAC
     5701 GGCGAGGATC TCGTCGTGAC CCATGGCGAT GCCTGCTTGC CGAATATCAT GGTGGAAAAT
     5761 GGCCGCTTTT CTGGATTCAT CGACTGTGGC CGGCTGGGTG TGGCGGACCG CTATCAGGAC
     5821 ATAGCGTTGG CTACCCGTGA TATTGCTGAA GAGCTTGGCG GCGAATGGGC TGACCGCTTC
     5881 CTCGTGCTTT ACGGTATCGC CGCTCCCGAT TCGCAGCGCA TCGCCTTCTA TCGCCTTCTT
     5941 GACGAGTTCT TCTGAGCGGG ACTCTGGGGT TCGAAATGAC CGACCAAGCG ACGCCCAACC
     6001 TGCCATCACG ATGGCCGCAA TAAAATATCT TTATTTTCAT TACATCTGTG TGTTGGTTTT
     6061 TTGTGTGAAT CGATAGCGAT AAGGATCCGC GTATGGTGCA CTCTCAGTAC AATCTGCTCT
     6121 GATGCCGCAT AGTTAAGCCA GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG
     6181 GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG
     6241 TGTCAGAGGT TTTCACCGTC ATCACCGAAA CGCGCGAGAC GAAAGGGCCT CGTGATACGC
     6301 CTATTTTTAT AGGTTAATGT CATGATAATA ATGGTTTCTT AGACGTCAGG TGGCACTTTT
     6361 CGGGGAAATG TGCGCGGAAC CCCTATTTGT TTATTTTTCT AAATACATTC AAATATGTAT
     6421 CCGCTCATGA GACAATAACC CTGATAAATG CTTCAATAAT ATTGAAAAAG GAAGAGTATG
     6481 AGTATTCAAC ATTTCCGTGT CGCCCTTATT CCCTTTTTTG CGGCATTTTG CCTTCCTGTT
     6541 TTTGCTCACC CAGAAACGCT GGTGAAAGTA AAAGATGCTG AAGATCAGTT GGGTGCACGA
     6601 GTGGGTTACA TCGAACTGGA TCTCAACAGC GGTAAGATCC TTGAGAGTTT TCGCCCCGAA
     6661 GAACGTTTTC CAATGATGAG CACTTTTAAA GTTCTGCTAT GTGGCGCGGT ATTATCCCGT
     6721 ATTGACGCCG GGCAAGAGCA ACTCGGTCGC CGCATACACT ATTCTCAGAA TGACTTGGTT
     6781 GAGTACTCAC CAGTCACAGA AAAGCATCTT ACGGATGGCA TGACAGTAAG AGAATTATGC
     6841 AGTGCTGCCA TAACCATGAG TGATAACACT GCGGCCAACT TACTTCTGAC AACGATCGGA
     6901 GGACCGAAGG AGCTAACCGC TTTTTTGCAC AACATGGGGG ATCATGTAAC TCGCCTTGAT
     6961 CGTTGGGAAC CGGAGCTGAA TGAAGCCATA CCAAACGACG AGCGTGACAC CACGATGCCT
     7021 GTAGCAATGG CAACAACGTT GCGCAAACTA TTAACTGGCG AACTACTTAC TCTAGCTTCC
     7081 CGGCAACAAT TAATAGACTG GATGGAGGCG GATAAAGTTG CAGGACCACT TCTGCGCTCG
     7141 GCCCTTCCGG CTGGCTGGTT TATTGCTGAT AAATCTGGAG CCGGTGAGCG TGGGTCTCGC
     7201 GGTATCATTG CAGCACTGGG GCCAGATGGT AAGCCCTCCC GTATCGTAGT TATCTACACG
     7261 ACGGGGAGTC AGGCAACTAT GGATGAACGA AATAGACAGA TCGCTGAGAT AGGTGCCTCA
     7321 CTGATTAAGC ATTGGTAACT GTCAGACCAA GTTTACTCAT ATATACTTTA GATTGATTTA
     7381 AAACTTCATT TTTAATTTAA AAGGATCTAG GTGAAGATCC TTTTTGATAA TCTCATGACC
     7441 AAAATCCCTT AACGTGAGTT TTCGTTCCAC TGAGCGTCAG ACCCCGTAGA AAAGATCAAA
     7501 GGATCTTCTT GAGATCCTTT TTTTCTGCGC GTAATCTGCT GCTTGCAAAC AAAAAAACCA
     7561 CCGCTACCAG CGGTGGTTTG TTTGCCGGAT CAAGAGCTAC CAACTCTTTT TCCGAAGGTA
     7621 ACTGGCTTCA GCAGAGCGCA GATACCAAAT ACTGTTCTTC TAGTGTAGCC GTAGTTAGGC
     7681 CACCACTTCA AGAACTCTGT AGCACCGCCT ACATACCTCG CTCTGCTAAT CCTGTTACCA
     7741 GTGGCTGCTG CCAGTGGCGA TAAGTCGTGT CTTACCGGGT TGGACTCAAG ACGATAGTTA
     7801 CCGGATAAGG CGCAGCGGTC GGGCTGAACG GGGGGTTCGT GCACACAGCC CAGCTTGGAG
     7861 CGAACGACCT ACACCGAACT GAGATACCTA CAGCGTGAGC TATGAGAAAG CGCCACGCTT
     7921 CCCGAAGGGA GAAAGGCGGA CAGGTATCCG GTAAGCGGCA GGGTCGGAAC AGGAGAGCGC
     7981 ACGAGGGAGC TTCCAGGGGG AAACGCCTGG TATCTTTATA GTCCTGTCGG GTTTCGCCAC
     8041 CTCTGACTTG AGCGTCGATT TTTGTGATGC TCGTCAGGGG GGCGGAGCCT ATGGAAAAAC
     8101 GCCAGCAACG CGGCCTTTTT ACGGTTCCTG GCCTTTTGCT GGCCTTTTGC TCACATGGCT
     8161 CGACAGATCT
//
