LOCUS       V044716                10560 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044716
VERSION     V044716
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..5240
                     /label="Pzp(NM_007376)"
                     /note="Pzp(NM_007376)"
                     /gene="Pzp"
     CDS             5247..5312
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5336..5924
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5990..6038
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6240..6668
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6682..7011
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7078..7872
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    8048..8169
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8218..8234)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8242..8258
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(8266..8296)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8311..8332
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8620..9208)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9379..10239)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(10240..10344)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgag gagaaaccag ctgcccacac
      781 cagcttttct tttactgttc ctgcttcttc ccagagatgc caccacagct actgcaaaac
      841 cacaatatgt ggtgctggtc ccgtcagagg tctattcagg agtccctgaa aaggcctgtg
      901 tcagcctcaa ccatgtgaat gagactgtga tgctcagctt aactctagag tatgcaatgc
      961 agcaaacgaa gctcctcaca gaccaggctg tggataagga ctccttctac tgcagcccct
     1021 tcacgatctc aggttcacct ttaccctaca cattcattac tgtggagata aaaggaccaa
     1081 cgcagcgctt cataaagaag aagtcaatac aaataataaa agctgagagc ccagtctttg
     1141 tccagacaga caaacccata tacaaaccag gacagatagt gaaattccga gttgtttctg
     1201 tggacatcag ttttcgcccg ttgaatgaaa cgttccctgt cgtttatatt gagactccca
     1261 agaggaaccg aatttttcaa tggcaaaata tccatctggc aggaggactc caccagctct
     1321 ctttcccact gtctgttgag ccagctctgg gtatctacaa ggttgtagtg cagaaggact
     1381 cagggaagaa aatagaacac tcctttgagg tgaaggaata cgttttaccc aaatttgagg
     1441 tgataataaa aatgcagaag actatggctt tcctggaaga ggaacttcct ataactgctt
     1501 gtggcgtata cacatatgga aagcctgttc ccggtctggt gacattgaga gtgtgcagaa
     1561 aatattcacg ataccgctcc acctgccata accaaaactc aatgagtatc tgtgaagaat
     1621 tcagccaaca ggcagatgat aaaggatgtt tcagacaagt tgtaaaaacc aaagtattcc
     1681 agctcagaca aaaaggccac gacatgaaga tagaggtgga agccaaaatc aaagaggaag
     1741 gaacaggaat agaactaact ggtattggat catgtgaaat agcaaatgcc ttaagcaaac
     1801 tgaaatttac taaagtaaat acaaattaca ggcctgggct acctttctct ggacaggttc
     1861 ttcttgttga tgagaagggt aaaccaatcc ccaacaaaaa cataacttcc gtcgtgtctc
     1921 cacttgggta cctgtccatt tttactacgg atgagcatgg cttggctaac atttccattg
     1981 acacttccaa cttcacagct ccgtttttga gagttgtggt cacctacaag cagaaccatg
     2041 tctgctatga taactggtgg cttgatgaat ttcacacaca ggcagaccat tctgcaactc
     2101 tagtcttttc tccaagccag agttatattc aacttgaact tgtttttggt actttggcct
     2161 gtgggcaaac tcaggagatt cggatacact acctcttgaa tgaagatatc atgaagaatg
     2221 aaaaagactt aaccttttac tatctgatca aagcaagggg aagcatcttc aacttaggaa
     2281 gccatgtgtt gtcgcttgaa caaggaaaca tgaaaggagt cttttccctc ccaattcaag
     2341 tggagccagg catggctccc gaggctcagc tgctcattta tgctatttta cctaatgaag
     2401 aacttgtcgc tgatgctcag aattttgaaa tcgagaagtg ttttgccaac aaggtaaatt
     2461 tgagtttccc atcagcacag agcctgccag cctctgacac ccacctgaag gtcaaagccg
     2521 cgcctctgtc cctctgtgcc ctcactgcag tagaccagag tgtgctgcta ctgaagcccg
     2581 aagccaagct ctcacctcag tcaatctaca atttgctgcc aggaaagact gtccagggtg
     2641 ccttctttgg tgtcccagtg tacaaagacc atgagaactg catcagcgga gaagacatca
     2701 ctcacaatgg gatcgtgtac acgccaaagc actctctggg cgataatgat gcacacagca
     2761 tttttcagtc tgtaggaata aatattttta ccaactccaa aatccacaaa ccacgctttt
     2821 gtcaagagtt tcaacactat ccggcaatgg gaggagtggc acctcaagcc ttagctgtgg
     2881 ctgcttcagg cccaggatcc agcttcagag caatgggcgt gccgatgatg gggttggact
     2941 actctgatga aattaatcag gtggtggaag taagagagac agtgcggaag tacttccctg
     3001 agacctggat ctgggacctg gtgccactgg acgtatccgg ggacggtgaa ttggcggtaa
     3061 aggtccccga caccatcact gagtggaagg ccagtgcatt ctgcttgtct ggaaccactg
     3121 gccttggcct ctcctccacc atctcccttc aagccttcca gcccttcttc ttggagctca
     3181 ctctcccata ctctgtggtg cgaggggaag cctttaccct caaagccacc gtgctcaatt
     3241 acatgtctca ctgcattcag atccgagtgg acctagagat ttctcctgat ttcctggcag
     3301 tcccagtggg gggccatgaa aactctcatt gcatctgtgg aaatgaaagg aaaaccgtgt
     3361 cctgggctgt gaccccaaag tcgctggggg aggtgaactt cacagctacc gcagaagcct
     3421 tgcagtctcc agaattgtgt ggcaataagt tgacagaagt gccagccctt gtacacaagg
     3481 acactgtggt gaagtccgta atagttgagc ctgaaggaat tgagaaggag caaacgtaca
     3541 acacactgtt atgcccacaa gatactgagt tacaagataa ttggtcactg gagcttccac
     3601 ccaatgtggt tgaaggatct gccagggcta cacattccgt tttgggtgat atactgggct
     3661 ctgcaatgca aaacctccag aatcttctcc agatgcccta tggctgtggg gaacaaaaca
     3721 tggtcctttt tgtccctaac atctatgttc tgaactatct gaacgagaca cagcagctga
     3781 cagaggcgat caagtccaaa gccattaact acctcatcag tgggtaccag aggcagctga
     3841 actatcagca cagtgatggc tcgtacagca cattcgggaa ccatggtggt ggcaacactc
     3901 cgggaaatac ttggctcact gcgtttgtgc tcaaggcctt tgctcaagct cagtcgcaca
     3961 tctttataga gaagacacac atcacaaatg ctttcaactg gctctccatg aaacaaaagg
     4021 agaatggctg tttccaacag tctggatatc tgcttaacaa tgcgatgaag ggtggtgtgg
     4081 atgatgaagt gactctctct gcctacatca ccattgctct gctggagatg cctctgcctg
     4141 tcacgcacag tgctgtgcga aatgctctat tttgcctgga aacggcctgg gcctccatct
     4201 cacagagcca ggaaagtcat gtctatacca aggcattgct ggcctatgcc ttcgccctgg
     4261 caggaaacaa ggccaagaga agcgaactgc ttgaatccct aaacaaagat gctgtgaagg
     4321 aagaggattc gctgcactgg caacgccctg gggatgttca gaaagtgaag gccttatctt
     4381 tctatcaacc tcgggcccct tctgccgaag tggagatgac agcttacgtg cttctcgcct
     4441 atctaacctc tgagtcttcc cggcccacac gggacctgtc ttcatcagac ctgtccacag
     4501 catccaagat tgtgaagtgg atcagcaagc aacaaaactc ccacgggggc ttctcctcca
     4561 ctcaggatac agtggtggct ctccaagccc tctccaaata tggagctgcc acttttacga
     4621 gaagtcagaa agaagtgttg gtcaccatcg agtcttcagg gaccttctct aagactttcc
     4681 atgttaacag tggcaatcgc ctgctgctgc aggaagtcag gctgccagat ctgcccggaa
     4741 attacgtcac caaagggtca ggatcaggat gtgtgtacct tcagacatct ctaaagtaca
     4801 acatcctccc agtggcagat ggaaaagcac ccttcgctct gcaagtcaac actctccccc
     4861 taaactttga caaggcagga gatcacagaa cattccagat tcgcatcaac gtaagctaca
     4921 ctggagaacg acccagctcc aacatggtca ttgttgatgt gaagatggta tcaggcttca
     4981 tacctatgaa gccatccgtg aaaaagctcc aagaccagcc taacattcag aggactgaag
     5041 tgaacaccaa ccatgttcta atctacattg aaaagctaac caatcaaacc ctcggtttct
     5101 ccttcgcggt ggaacaagac atcccagtaa agaacttaaa accagccccc ataaaagtct
     5161 atgattatta tgagacagat gaattcaccg ttgaagaata cagtgcccct ttcagtgatg
     5221 gctctgaaca aggaaatgct ggatccgact acaaagacca tgacggtgat tataaagatc
     5281 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     5341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5461 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5521 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5581 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5641 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5701 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     5761 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5821 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     5881 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     5941 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     6001 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6061 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6121 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6181 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     6241 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6301 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6361 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     6421 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6481 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6541 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6601 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6661 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     6961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     7021 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     7081 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     7141 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     7201 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     7261 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     7321 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     7381 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     7441 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     7501 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7561 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     7621 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7681 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7741 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7801 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7861 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     7921 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7981 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     8041 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8101 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8161 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     8221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     8281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     8341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     8401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     8461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     8521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     8701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     8761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     8821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     8881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     8941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     9001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     9061 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     9121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     9181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     9241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     9301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     9361 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     9421 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     9481 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     9541 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     9601 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     9661 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     9721 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     9781 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     9841 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     9901 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     9961 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    10021 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    10081 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    10141 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    10201 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    10261 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    10321 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
    10381 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
    10441 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
    10501 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
