LOCUS       40924_205        8799 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector AF286077, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8799)
  AUTHORS   Wang RF, Mullins JI.
  TITLE     Mammalian cell/vaccinia virus expression vectors with increased 
            stability of retroviral sequences in Escherichia coli: production of
            feline immunodeficiency virus envelope protein
  JOURNAL   Gene 153 (2), 197-202 (1995)
  PUBMED    7875588
REFERENCE   2  (bases 1 to 8799)
  AUTHORS   Mullins JI, Hanley T, Kafsack BFC., Rowen L.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2000) Microbiology, University of Washington, HSB 
            K-455, Seattle, Washington 98195, USA
REFERENCE   3  (bases 1 to 8799)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8799)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1995"; volume: "153"; issue: "2"; pages: "197-202"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JUL-2000) Microbiology, University of Washington, HSB K-455, 
            Seattle, Washington 98195, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            46..500
                     /pseudo
                     /gene="thymidine kinase"
                     /label=thymidine kinase
                     /note="derived from vaccinia; 5' end"
     promoter        526..1140
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     intron          1181..1274
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     promoter        1302..1320
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1413..1869
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1917..1934
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    1960..2007
                     /label=similar to tpa 3' end
                     /note="similar to tpa 3' end"
     polyA_signal    2031..2255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      2301..2331
                     /label=synthetic
                     /note="synthetic"
                     /regulatory_class="polyA_signal_sequence"
     terminator      2396..2443
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      2474..2747
                     /label=derived from P7.5 vaccinia
                     /note="derived from P7.5 vaccinia"
                     /regulatory_class="promoter"
     misc_feature    2777..3143
                     /label=murine beta-globin transcriptional regulation unit
                     /note="murine beta-globin transcriptional regulation unit"
     CDS             3244..3804
                     /label=DHFR
                     /note="mouse dihydrofolate reductase"
     polyA_signal    3900..4107
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4142..4507
                     /label=murine beta-globin transcriptional regulation unit
                     /note="murine beta-globin transcriptional regulation unit"
     CDS             4528..5319
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5496..5630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5638..6265
                     /label=vaccinia thymidine kinase gene cluster 3' end
                     /note="vaccinia thymidine kinase gene cluster 3' end"
     gene            5638..5911
                     /pseudo
                     /gene="thymidine kinase"
                     /label=thymidine kinase
                     /note="derived from vaccinia; 3' end"
     misc_feature    6320..6449
                     /label=similar to pUC19
                     /note="similar to pUC19"
     rep_origin      6489..7033
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(7195..8052)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8053..8157)
                     /label=AmpR promoter
     rep_origin      complement(8183..8638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 accaattgac aaaattcaca gactttcaag attttaaaaa actgtttaac aaggtcccta
       61 ttgttacaga tggaagggtc aaacttaata aaggatattt gttcgacttt gtgattagtt
      121 tgatgcgatt caaaaaagaa tcctctctag ctaccaccgc aatagatcct gttagataca
      181 tagatcctcg tcgcaatatc gcattttcta acgtgatgga tatattaaag tcgaataaag
      241 tgaacaataa ttaattcttt attgtcatca tgaacggcgg acatattcag ttgataatcg
      301 gccccatgtt ttcaggtaaa agtacagaat taattagacg agttagacgt tatcaaatag
      361 ctcaatataa atgcgtgact ataaaatatt ctaacgataa tagatacgga acgggactat
      421 ggacgcatga taagaataat tttgaagcat tggaagcaac taaactatgt gatgtcttgg
      481 aatcaattac agatttctcc gtgataggta tcgcttctgt ggaatgtgtg tcagttaggg
      541 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      601 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      661 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
      721 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
      781 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
      841 ctaggctttt gcaaaaagct cctccgatcg aggggctcgc atctctcctt cacgcgcccg
      901 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
      961 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
     1021 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
     1081 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
     1141 caagctgcct cgaggaactg aaaaaccaga aagttaactg gtaagtttag tctttttgtc
     1201 ttttatttca ggtcccggat ccggtggtgc aaatcaaaga actgctcctc agtggatgtt
     1261 gcctttactt ctaggcctgt acggaagtgt tacttctgct ctaatacgac tcactatagg
     1321 aagcttcgta cgaagaagga gatcttggaa taaggccggt gtgcgtttgt ctatatgtta
     1381 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     1441 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     1501 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     1561 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     1621 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     1681 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     1741 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
     1801 tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     1861 aaacacgatt taaatatggc tagccatatg tacgtaccgc gggccctcta gaattccatc
     1921 accatcacca tcacggggcg cccgcatagg gatccgttaa cggttaccaa ctacctagac
     1981 tggattcgtg acaacatgcg gccgtgatat ctacgtatga tcagcctcga ctgtgccttc
     2041 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2101 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2161 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2221 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2281 gggctcgaca gcaaacgcgt aataaaaaaa aaaaaaaaaa aaaaaaaaaa acgatccggc
     2341 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     2401 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     2461 atccggagtt aactcgacat atactatata gtaataccaa tactcaagac tacgaaactg
     2521 atacaatctc ttatcatgtg ggtaatgttc tcgatgtcga atagccatat gccggtagtt
     2581 gcgatataca taaactgatc actaattcca aacccacccg ctttttatag taagtttttc
     2641 acccataaat aataaataca ataattaatt tctcgtaaaa gtagaaaata tattctaatt
     2701 tattgcacgg taaggaagta gaatcataaa gaacagtgac ggatcgatcc cccaagcgct
     2761 ggatctcccg atccccagct ttgcttctca atttcttatt tgcataatga gaaaaaaagg
     2821 aaaattaatt ttaacaccaa ttcagtagtt gattgagcaa atgcgttgcc aaaaaggatg
     2881 ctttagagac agtgttctct gcacagataa ggacaaacat tattcagagg gagtacccag
     2941 agctgagact cctaagccag tgagtggcac agcattctag ggagaaatat gcttgtcatc
     3001 accgaagcct gattccgtag agccacacct tggtaagggc caatctgctc acacaggata
     3061 gagagggcag gagccagggc agagcatata aggtgaggta ggatcagttg ctcctcacat
     3121 ttgcttctga catagttgtg ttgggagctt ggatagcttg gacagctcag ggctgcgatt
     3181 tcgcgccaaa cttgacggca atcctagcgt gaaggctggt aggattttat ccccgctgcc
     3241 atcatggttc gaccattgaa ctgcatcgtc gccgtgtccc aaaatatggg gattggcaag
     3301 aacggagacc taccctggcc tccgctcagg aacgagttca agtacttcca aagaatgacc
     3361 acaacctctt cagtggaagg taaacagaat ctggtgatta tgggtaggaa aacctggttc
     3421 tccattcctg agaagaatcg acctttaaag gacagaatta atatagttct cagtagagaa
     3481 ctcaaagaac caccacgagg agctcatttt cttgccaaaa gtttggatga tgccttaaga
     3541 cttattgaac aaccggaatt ggcaagtaaa gtagacatgg tttggatagt cggaggcagt
     3601 tctgtttacc aggaagccat gaatcaacca ggccacctta gactctttgt gacaaggatc
     3661 atgcaggaat ttgaaagtga cacgtttttc ccagaaattg atttggggaa atataaactt
     3721 ctcccagaat acccaggcgt cctctctgag gtccaggagg aaaaaggcat caagtataag
     3781 tttgaagtct acgagaagaa agactaacag gaagatgctt tcaagttctc tgctcccctc
     3841 ctaaagctat gcatttttat aagaccatgg gacttttgct ggctttagat cagcctcgac
     3901 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3961 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4021 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4081 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggaaccag ctggggctcg
     4141 agctttgctt ctcaatttct tatttgcata atgagaaaaa aaggaaaatt aattttaaca
     4201 ccaattcagt agttgattga gcaaatgcgt tgccaaaaag gatgctttag agacagtgtt
     4261 ctctgcacag ataaggacaa acattattca gagggagtac ccagagctga gactcctaag
     4321 ccagtgagtg gcacagcatt ctagggagaa atatgcttgt catcaccgaa gcctgattcc
     4381 gtagagccac accttggtaa gggccaatct gctcacacag gatagagagg gcaggagcca
     4441 gggcagagca tataaggtga ggtaggatca gttgctcctc acatttgctt ctgacatagt
     4501 tgtgttggga gcttggatcg atccaccatg gttgaacaag atggattgca cgcaggttct
     4561 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4621 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4681 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4741 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4801 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4861 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4921 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4981 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5041 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5101 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5161 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5221 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5281 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5341 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5401 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5461 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5521 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5581 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gcggccctcc
     5641 ggatcgagat caattctgtg agcgtatggc aaacgaagga aaaatagtta tagtagccgc
     5701 actcgatggg acatttcaac gtaaaccgtt taataatatt ttgaatctta ttccattatc
     5761 tgaaatggtg gtaaaactaa ctgctgtgtg tatgaaatgc tttaaggagg cttccttttc
     5821 taaacgattg ggtgaggaaa ccgagataga aataatagga ggtaatgata tgtatcaatc
     5881 ggtgtgtaga aagtgttaca tcgactcata atattatatt ttttatctaa aaaactaaaa
     5941 ataaacattg attaaatttt aatataatac ttaaaaatgg atgttgtgtc gttagataaa
     6001 ccgtttatgt attttgagga aattgataat gagttagatt acgaaccaga aagtgcaaat
     6061 gaggtcgcaa aaaaactgcc gtatcaagga cagttaaaac tattactagg agaattattt
     6121 tttcttagta agttacagcg acacggtata ttagatggtg ccaccgtagt gtatatagga
     6181 tctgctcccg gtacacatat acgttatttg agagatcatt tctataattt aggagtgatc
     6241 atcaaatgga tgctaattga cggccctgca ttaatgaatc ggccaacgcg cggggagagg
     6301 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6361 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6421 aggggataac gcaggaaaga acatgtgaga tcctagaaat attttatctg attaataaga
     6481 tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc
     6541 gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     6601 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     6661 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     6721 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     6781 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     6841 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     6901 caggaacagg agagcgcacg agggagccgc caggggaaac gcctggtatc tttatagtcc
     6961 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     7021 gagcctatgg aaaaacggct ttgccggatc tgacgctcag tggaacgaaa actcacgtta
     7081 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7141 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7201 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7261 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7321 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7381 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7441 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7501 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7561 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7621 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7681 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7741 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7801 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7861 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7921 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7981 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8041 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8101 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8161 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     8221 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     8281 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     8341 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     8401 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     8461 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     8521 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     8581 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     8641 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     8701 ttacgccagc tagactttct gttattatta ttgatccaat caaaaaataa attagaagcc
     8761 gtgggtcatt gttatgaatc tctttcagag gaatacaga
//
