LOCUS       V021752                 8681 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021752
VERSION     V021752
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"
     CDS             683..3328
                     /label="PML(NM_033238)"
                     /note="PML(NM_033238)"
                     /gene="PML"
     CDS             3368..3433
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3457..4045
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4111..4159
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4361..4789
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4803..5132
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5199..5993
                     /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    6169..6290
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6339..6355)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6363..6379
                     /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(6387..6417)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6432..6453
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6741..7329)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7500..8360)
                     /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(8361..8465)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggagcc tgcacccgcc cgatctccga ggccccagca
      721 ggaccccgcc cggccccagg agcccaccat gcctcccccc gagaccccct ctgaaggccg
      781 ccagcccagc cccagcccca gccctacaga gcgagccccc gcttcggagg aggagttcca
      841 gtttctgcgc tgccagcaat gccaggcgga agccaagtgc ccgaagctgc tgccttgtct
      901 gcacacgctg tgctcaggat gcctggaggc gtcgggcatg cagtgcccca tctgccaggc
      961 gccctggccc ctaggtgcag acacacccgc cctggataac gtctttttcg agagtctgca
     1021 gcggcgcctg tcggtgtacc ggcagattgt ggatgcgcag gctgtgtgca cccgctgcaa
     1081 agagtcggcc gacttctggt gctttgagtg cgagcagctc ctctgcgcca agtgcttcga
     1141 ggcacaccag tggttcctca agcacgaggc ccggccccta gcagagctgc gcaaccagtc
     1201 ggtgcgtgag ttcctggacg gcacccgcaa gaccaacaac atcttctgct ccaaccccaa
     1261 ccaccgcacc cctacgctga ccagcatcta ctgccgagga tgttccaagc cgctgtgctg
     1321 ctcgtgcgcg ctccttgaca gcagccacag tgagctcaag tgcgacatca gcgcagagat
     1381 ccagcagcga caggaggagc tggacgccat gacgcaggcg ctgcaggagc aggatagtgc
     1441 ctttggcgcg gttcacgcgc agatgcacgc ggccgtcggc cagctgggcc gcgcgcgtgc
     1501 cgagaccgag gagctgatcc gcgagcgcgt gcgccaggtg gtagctcacg tgcgggctca
     1561 ggagcgcgag ctgctggagg ctgtggacgc gcggtaccag cgcgactacg aggagatggc
     1621 cagtcggctg ggccgcctgg atgctgtgct gcagcgcatc cgcacgggca gcgcgctggt
     1681 gcagaggatg aagtgctacg cctcggacca ggaggtgctg gacatgcacg gtttcctgcg
     1741 ccaggcgctc tgccgcctgc gccaggagga gccccagagc ctgcaagctg ccgtgcgcac
     1801 cgatggcttc gacgagttca aggtgcgcct gcaggacctc agctcttgca tcacccaggg
     1861 gaaagatgca gctgtatcca agaaagccag cccagaggct gccagcactc ccagggaccc
     1921 tattgacgtt gacctgcccg aggaggcaga gagagtgaag gcccaggttc aggccctggg
     1981 gctggctgaa gcccagccta tggctgtggt acagtcagtg cccggggcac accccgtgcc
     2041 agtgtacgcc ttctccatca aaggcccttc ctatggagag gatgtctcca atacaacgac
     2101 agcccagaag aggaagtgca gccagaccca gtgccccagg aaggtcatca agatggagtc
     2161 tgaggagggg aaggaggcaa ggttggctcg gagctccccg gagcagccca ggcccagcac
     2221 ctccaaggca gtctcaccac cccacctgga tggaccgcct agccccagga gccccgtcat
     2281 aggaagtgag gtcttcctgc ccaacagcaa ccacgtggcc agtggcgccg gggaggcaga
     2341 ggaacgcgtt gtggtgatca gcagctcgga agactcagat gccgaaaact cgtcctcccg
     2401 agagctggat gacagcagca gtgagtccag tgacctccag ctggaaggcc ccagcaccct
     2461 cagggtcctg gacgagaacc ttgctgaccc ccaagcagaa gacagacctc tggttttctt
     2521 tgacctcaag attgacaatg aaacccagaa gattagccag ctggctgcgg tgaaccggga
     2581 aagcaagttc cgcgtggtca tccagcctga agccttcttc agcatctact ccaaggccgt
     2641 gtccctggag gtggggctgc agcacttcct cagctttctg agctccatgc gccgccctat
     2701 cttggcctgc tacaagctgt gggggcctgg cctcccaaac ttcttccggg ccctggagga
     2761 cattaacagg ctgtgggaat tccaggaggc catctcgggc ttcctggctg ccctgcctct
     2821 catccgggag cgtgtgcccg gggccagcag cttcaaactc aagaacctgg cccagaccta
     2881 cctggcgaga aacatgagcg agcgcagcgc catggctgcc gtgctggcca tgcgtgacct
     2941 gtgccgcctc ctcgaggtct ccccgggccc ccagctggcc cagcatgtct accccttcag
     3001 tagcctgcag tgctttgcct ccctgcagcc cctggtgcag gcagctgtgc tgccccgggc
     3061 tgaggcccgc ctcctggccc tacacaacgt gagcttcatg gagctgctga gtgcacaccg
     3121 ccgtgaccgg caggggggcc tgaagaagta cagccgctat ctaagcctgc agaccaccac
     3181 gttgccccct gcccagcctg ctttcaacct gcaggctctg ggcacctact ttgaaggcct
     3241 gttggagggt ccggcgctgg cacgggcaga aggagtctcc accccacttg ctggccgtgg
     3301 cttggcagag agggcctccc agcagagcga attctgcaga tatccagcac agtggcggcc
     3361 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3421 tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt
     3481 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3541 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3601 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3661 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3721 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3781 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3841 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     3901 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3961 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4021 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg
     4081 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4141 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4201 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4261 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4321 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4381 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4441 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4501 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4561 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4621 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4681 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     4741 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4801 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4861 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4921 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4981 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5041 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5101 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5161 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5221 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5281 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5341 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5401 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5461 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5521 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5581 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5641 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5701 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5761 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5821 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5881 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5941 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6001 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     6061 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     6121 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     6181 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6241 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6301 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6361 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6421 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6481 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6541 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6601 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6661 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6901 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7141 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7201 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7261 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7321 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7381 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7441 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7501 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7561 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7621 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7681 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7741 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7801 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7861 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7921 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7981 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8041 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8101 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8161 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8221 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8281 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8341 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8401 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8461 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8521 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     8581 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     8641 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
