LOCUS       V041651                 8554 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041651
VERSION     V041651
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             706..3210
                     /label="TRIM28(NM_005762)"
                     /note="TRIM28(NM_005762)"
                     /gene="TRIM28"
     CDS             3217..3306
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3330..3918
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3984..4032
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4234..4662
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4676..5005
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5072..5866
                     /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    6042..6163
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6212..6228)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6236..6252
                     /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(6260..6290)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6305..6326
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6614..7202)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7373..8233)
                     /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(8234..8338)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc ggcctccgcg
      721 gcggcagcct cggcagcagc ggcctcggcc gcctctggca gcccgggccc gggcgagggc
      781 tccgctggcg gcgaaaagcg ctccaccgcc ccttcggccg cagcctcggc ctctgcctca
      841 gccgcggcgt cgtcgcccgc ggggggcggc gccgaggcgc tggagctgct ggagcactgc
      901 ggcgtgtgca gagagcgcct gcgacccgag agggagcccc gcctgctgcc ctgtttgcac
      961 tcggcctgta gtgcctgctt agggcccgcg gcccccgccg ccgccaacag ctcgggggac
     1021 ggcggggcgg cgggcgacgg caccgtggtg gactgtcccg tgtgcaagca acagtgcttc
     1081 tccaaagaca tcgtggagaa ttatttcatg cgtgatagtg gcagcaaggc tgccaccgac
     1141 gcccaggatg cgaaccagtg ctgcactagc tgtgaggata atgccccagc caccagctac
     1201 tgtgtggagt gctcggagcc tctgtgtgag acctgtgtag aggcgcacca gcgggtgaag
     1261 tacaccaagg accatactgt gcgctctact gggccagcca agtctcggga tggtgaacgt
     1321 actgtctatt gcaacgtaca caagcatgaa ccccttgtgc tgttttgtga gagctgtgat
     1381 actctcacct gccgagactg ccagctcaat gcccacaagg accaccagta ccagttctta
     1441 gaggatgcag tgaggaacca gcgcaagctc ctggcctcac tggtgaagcg ccttggggac
     1501 aaacatgcaa cattgcagaa gagcaccaag gaggttcgca gctcaatccg ccaggtgtct
     1561 gacgtacaga agcgtgtgca agtggatgtc aagatggcca tcctgcagat catgaaggag
     1621 ctgaataagc ggggccgtgt gctggtcaat gatgcccaga aggtgactga ggggcagcag
     1681 gagcgcctgg agcggcagca ctggaccatg accaagatcc agaagcacca ggagcacatt
     1741 ctgcgctttg cctcttgggc tctggagagt gacaacaaca cagccctttt gctttctaag
     1801 aagttgatct acttccagct gcaccgggcc ctcaagatga ttgtggatcc cgtggagcca
     1861 catggcgaga tgaagtttca gtgggacctc aatgcctgga ccaagagtgc cgaggccttt
     1921 ggcaagattg tggcagagcg tcctggcact aactcaacag gccctgcacc catggcccct
     1981 ccaagagccc cagggcccct gagcaagcag ggctctggca gcagccagcc catggaggtg
     2041 caggaaggct atggctttgg gtcaggagat gatccctact caagtgcaga gccccatgtg
     2101 tcaggtgtga aacggtcccg ctcaggtgag ggcgaggtga gcggccttat gcgcaaggtg
     2161 ccacgagtga gccttgaacg cctggacctg gacctcacag ctgacagcca gccacccgtc
     2221 ttcaaggtct tcccaggcag taccactgag gactacaacc ttattgttat tgaacgtggc
     2281 gctgccgctg cagctaccgg ccagccaggg actgcgcctg caggaacccc tggtgcccca
     2341 cccctggctg gcatggccat tgtcaaggag gaggagacgg aggctgccat tggagcccct
     2401 cctactgcca ctgagggccc tgagaccaaa cctgtgctta tggctcttgc ggagggtcct
     2461 ggtgctgagg gtccccgcct ggcctcacct agtggcagca ccagctcagg gctggaggtg
     2521 gtggctcctg agggtacctc agccccaggt ggtggcccgg gaaccctgga tgacagtgcc
     2581 accatttgcc gtgtctgcca gaagccaggc gatctggtta tgtgcaacca gtgtgagttt
     2641 tgtttccacc tggactgtca cctgccggcc ctgcaggatg taccagggga ggagtggagc
     2701 tgctcactct gccatgtgct ccctgacctg aaggaggagg atggcagcct cagcctggat
     2761 ggtgcagaca gcactggcgt ggtggccaag ctctcaccag ccaaccagcg gaaatgtgag
     2821 cgtgtactgc tggccctatt ctgtcacgaa ccctgccgcc ccctgcatca gctggctacc
     2881 gactccacct tctccctgga ccagcccggt ggcaccctgg atctgaccct gatccgtgcc
     2941 cgcctccagg agaagttgtc acctccctac agctccccac aggagtttgc ccaggatgtg
     3001 ggccgcatgt tcaagcaatt caacaagtta actgaggaca aggcagacgt gcagtccatc
     3061 atcggcctgc agcgcttctt cgagacgcgc atgaacgagg ccttcggtga caccaagttc
     3121 tctgctgtgc tggtggagcc cccgccgatg agcctgcctg gtgctggcct gagttcccag
     3181 gagctgtctg gtggccctgg tgatggcccc ctcgaggaac aaaaactaat atcagaggaa
     3241 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag
     3301 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     3361 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3421 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3481 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3541 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3601 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3661 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3721 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3781 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3841 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3901 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     3961 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     4021 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4081 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4141 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     4201 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4261 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4321 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4381 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4441 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4501 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4561 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     4621 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4681 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4741 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4801 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4861 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4921 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4981 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5041 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5101 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5161 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5221 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5281 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5341 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5401 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5461 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5521 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5581 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5641 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5701 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5761 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5821 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5881 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5941 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6001 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6061 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6181 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6301 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6361 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6781 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7021 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8401 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     8461 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     8521 cttgaccgac aattgcatga agaatctgct tagg
//
