LOCUS       V020378                 8219 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020378
VERSION     V020378
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..2899
                     /label="SUZ12(NM_015355)"
                     /note="SUZ12(NM_015355)"
                     /gene="SUZ12"
     CDS             2906..2971
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2995..3583
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3649..3697
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3899..4327
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4341..4670
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4737..5531
                     /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    5707..5828
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5877..5893)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5901..5917
                     /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(5925..5955)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5970..5991
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6279..6867)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7038..7898)
                     /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(7899..8003)
                     /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 ccatggcgcc tcagaagcac ggcggtgggg gagggggcgg
      721 ctcggggccc agcgcggggt ccgggggagg cggcttcggg ggttcggcgg cggtggcggc
      781 ggcgacggct tcgggcggca aatccggcgg cgggagctgt ggagggggtg gcagttactc
      841 ggcctcctcc tcctcctccg cggcggcagc ggcgggggct gcggtgttac cggtgaagaa
      901 gccgaaaatg gagcacgtcc aggctgacca cgagcttttc ctccaggcct ttgagaagcc
      961 aacacagatc tatagatttc ttcgaactcg gaatctcata gcaccaatat ttttgcacag
     1021 aactcttact tacatgtctc atcgaaactc cagaacaaac atcaaaagga aaacatttaa
     1081 agttgatgat atgttatcaa aagtagagaa aatgaaagga gagcaagaat ctcatagctt
     1141 gtcagctcat ttgcagctta cgtttactgg tttcttccac aaaaatgata agccatcacc
     1201 aaactcagaa aatgaacaaa attctgttac cctggaagtc ctgcttgtga aagtttgcca
     1261 caaaaaaaga aaggatgtaa gttgtccaat aaggcaagtt cccacaggta aaaagcaggt
     1321 gcctttgaat cctgacctca atcaaacaaa acccggaaat ttcccgtccc ttgcagtttc
     1381 cagtaatgaa tttgaaccta gtaacagcca tatggtgaag tcttactcgt tgctatttag
     1441 agtgactcgt ccaggaagaa gagagtttaa tggaatgatt aatggagaaa ccaatgaaaa
     1501 tattgatgtc aatgaagagc ttccagccag aagaaaacga aatcgtgagg atggggaaaa
     1561 gacatttgtt gcacaaatga cagtatttga taaaaacagg cgcttacagc ttttagatgg
     1621 ggaatatgaa gtagccatgc aggaaatgga agaatgtcca ataagcaaga aaagagcaac
     1681 atgggagact attcttgatg ggaagaggct gcctccattc gaaacatttt ctcagggacc
     1741 tacgttgcag ttcactcttc gttggacagg agagaccaat gataaatcta cggctcctat
     1801 tgccaaacct cttgccacta gaaattcaga gagtctccat caggaaaaca agcctggttc
     1861 agttaaacct actcaaacta ttgctgttaa agaatcattg actacagatc tacaaacaag
     1921 aaaagaaaag gatactccaa atgaaaaccg acaaaaatta agaatatttt atcagtttct
     1981 ctataacaac aatacaaggc aacaaactga agcaagagat gacctgcatt gcccttggtg
     2041 tactctgaac tgccgcaaac tttatagttt actcaagcat cttaaactct gccatagcag
     2101 atttatcttc aactatgttt atcatccaaa aggtgctagg atagatgttt ctatcaatga
     2161 gtgttatgat ggctcctatg caggaaatcc tcaggatatt catcgccaac ctggatttgc
     2221 ttttagtcgc aacggaccag ttaagagaac acctatcaca catattcttg tgtgcaggcc
     2281 aaaacgaaca aaagcaagca tgtctgaatt tcttgaatct gaagatgggg aagtagaaca
     2341 gcaaagaaca tatagtagtg gccacaatcg tctgtatttc catagtgata cctgcttacc
     2401 tctccgtcca caagaaatgg aagtagatag tgaagatgaa aaggatcctg aatggctaag
     2461 agaaaaaacc attacacaaa ttgaagagtt ttctgatgtt aatgaaggag agaaagaagt
     2521 gatgaaactc tggaatctcc atgtcatgaa gcatgggttt attgctgaca atcaaatgaa
     2581 tcatgcctgt atgctgtttg tagaaaatta tggacagaaa ataattaaga agaatttatg
     2641 tcgaaacttc atgcttcatc tagtcagcat gcatgacttt aatcttatta gcataatgtc
     2701 aatagataaa gctgttacca agctccgtga aatgcagcaa aaattagaaa agggggaatc
     2761 tgcttcccct gcaaacgaag aaataactga agaacaaaat gggacagcaa atggatttag
     2821 tgaaattaac tcaaaagaga aagctttgga aacagatagt gtctcagggg tttcaaaaca
     2881 gagcaaaaaa caaaaactcc tcgaggacta caaagaccat gacggtgatt ataaagatca
     2941 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     3001 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3061 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3121 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3181 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3241 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3301 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3361 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     3421 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3481 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3541 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     3601 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3661 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3721 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     3781 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     3841 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     3901 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3961 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4021 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     4081 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4141 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4201 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4261 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4321 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4501 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4561 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4621 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4681 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4741 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4801 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4861 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4921 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4981 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5041 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5101 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5161 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5221 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5281 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5341 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5401 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5461 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5521 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     5581 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5641 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5701 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5761 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5821 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5881 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5941 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6001 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6061 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     6481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     8041 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     8101 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     8161 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
