LOCUS       V020117                 8786 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020117
VERSION     V020117
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..3466
                     /label="RB1(NM_000321)"
                     /note="RB1(NM_000321)"
                     /gene="RB1"
     CDS             3473..3538
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3562..4150
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4216..4264
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4466..4894
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4908..5237
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5304..6098
                     /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    6274..6395
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6444..6460)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6468..6484
                     /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(6492..6522)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6537..6558
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6846..7434)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7605..8465)
                     /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(8466..8570)
                     /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 ccatgccgcc caaaaccccc cgaaaaacgg ccgccaccgc
      721 cgccgctgcc gccgcggaac ccccggcacc gccgccgccg ccccctcctg aggaggaccc
      781 agagcaggac agcggcccgg aggacctgcc tctcgtcagg cttgagtttg aagaaacaga
      841 agaacctgat tttactgcat tatgtcagaa attaaagata ccagatcatg tcagagagag
      901 agcttggtta acttgggaga aagtttcatc tgtggatgga gtattgggag gttatattca
      961 aaagaaaaag gaactgtggg gaatctgtat ctttattgca gcagttgacc tagatgagat
     1021 gtcgttcact tttactgagc tacagaaaaa catagaaatc agtgtccata aattctttaa
     1081 cttactaaaa gaaattgata ccagtaccaa agttgataat gctatgtcaa gactgttgaa
     1141 gaagtatgat gtattgtttg cactcttcag caaattggaa aggacatgtg aacttatata
     1201 tttgacacaa cccagcagtt cgatatctac tgaaataaat tctgcattgg tgctaaaagt
     1261 ttcttggatc acatttttat tagctaaagg ggaagtatta caaatggaag atgatctggt
     1321 gatttcattt cagttaatgc tatgtgtcct tgactatttt attaaactct cacctcccat
     1381 gttgctcaaa gaaccatata aaacagctgt tatacccatt aatggttcac ctcgaacacc
     1441 caggcgaggt cagaacagga gtgcacggat agcaaaacaa ctagaaaatg atacaagaat
     1501 tattgaagtt ctctgtaaag aacatgaatg taatatagat gaggtgaaaa atgtttattt
     1561 caaaaatttt atacctttta tgaattctct tggacttgta acatctaatg gacttccaga
     1621 ggttgaaaat ctttctaaac gatacgaaga aatttatctt aaaaataaag atctagatgc
     1681 aagattattt ttggatcatg ataaaactct tcagactgat tctatagaca gttttgaaac
     1741 acagagaaca ccacgaaaaa gtaaccttga tgaagaggtg aatgtaattc ctccacacac
     1801 tccagttagg actgttatga acactatcca acaattaatg atgattttaa attcagcaag
     1861 tgatcaacct tcagaaaatc tgatttccta ttttaacaac tgcacagtga atccaaaaga
     1921 aagtatactg aaaagagtga aggatatagg atacatcttt aaagagaaat ttgctaaagc
     1981 tgtgggacag ggttgtgtcg aaattggatc acagcgatac aaacttggag ttcgcttgta
     2041 ttaccgagta atggaatcca tgcttaaatc agaagaagaa cgattatcca ttcaaaattt
     2101 tagcaaactt ctgaatgaca acatttttca tatgtcttta ttggcgtgcg ctcttgaggt
     2161 tgtaatggcc acatatagca gaagtacatc tcagaatctt gattctggaa cagatttgtc
     2221 tttcccatgg attctgaatg tgcttaattt aaaagccttt gatttttaca aagtgatcga
     2281 aagttttatc aaagcagaag gcaacttgac aagagaaatg ataaaacatt tagaacgatg
     2341 tgaacatcga atcatggaat cccttgcatg gctctcagat tcacctttat ttgatcttat
     2401 taaacaatca aaggaccgag aaggaccaac tgatcacctt gaatctgctt gtcctcttaa
     2461 tcttcctctc cagaataatc acactgcagc agatatgtat ctttctcctg taagatctcc
     2521 aaagaaaaaa ggttcaacta cgcgtgtaaa ttctactgca aatgcagaga cacaagcaac
     2581 ctcagccttc cagacccaga agccattgaa atctacctct ctttcactgt tttataaaaa
     2641 agtgtatcgg ctagcctatc tccggctaaa tacactttgt gaacgccttc tgtctgagca
     2701 cccagaatta gaacatatca tctggaccct tttccagcac accctgcaga atgagtatga
     2761 actcatgaga gacaggcatt tggaccaaat tatgatgtgt tccatgtatg gcatatgcaa
     2821 agtgaagaat atagacctta aattcaaaat cattgtaaca gcatacaagg atcttcctca
     2881 tgctgttcag gagacattca aacgtgtttt gatcaaagaa gaggagtatg attctattat
     2941 agtattctat aactcggtct tcatgcagag actgaaaaca aatattttgc agtatgcttc
     3001 caccaggccc cctaccttgt caccaatacc tcacattcct cgaagccctt acaagtttcc
     3061 tagttcaccc ttacggattc ctggagggaa catctatatt tcacccctga agagtccata
     3121 taaaatttca gaaggtctgc caacaccaac aaaaatgact ccaagatcaa gaatcttagt
     3181 atcaattggt gaatcattcg ggacttctga gaagttccag aaaataaatc agatggtatg
     3241 taacagcgac cgtgtgctca aaagaagtgc tgaaggaagc aaccctccta aaccactgaa
     3301 aaaactacgc tttgatattg aaggatcaga tgaagcagat ggaagtaaac atctcccagg
     3361 agagtccaaa tttcagcaga aactggcaga aatgacttct actcgaacac gaatgcaaaa
     3421 gcagaaaatg aatgatagca tggatacctc aaacaaggaa gagaaactcg aggactacaa
     3481 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     3541 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact
     3601 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3661 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3721 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3781 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3841 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3901 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     3961 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4021 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4081 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4141 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4201 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     4261 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4321 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4381 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     4441 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4621 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4681 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4741 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4801 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     4861 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4921 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4981 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5041 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5161 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5221 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5281 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5341 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5401 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5461 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5521 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5581 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5641 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5701 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5761 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5821 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5881 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5941 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6001 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6061 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     6121 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6181 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6241 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     6301 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6361 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     6421 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6481 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6541 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6601 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6661 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6961 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7021 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7261 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     7321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7501 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7561 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7621 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7681 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7741 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7801 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7861 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7921 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7981 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8041 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8101 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8161 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8221 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8281 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8341 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8401 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8461 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8521 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8581 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     8641 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     8701 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     8761 acaattgcat gaagaatctg cttagg
//
