LOCUS       V032561                 7393 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032561
VERSION     V032561
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..2883
                     /label="ZNF549(NM_001199295)"
                     /note="ZNF549(NM_001199295)"
                     /gene="ZNF549"
     CDS             2890..2955
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2993..3217
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3263..3691
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3705..4034
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4101..4895
                     /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    5069..5190
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5239..5255)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5263..5279
                     /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(5287..5317)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5332..5353
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5641..6226)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6397..7257)
                     /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(7258..7362)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggccg aggcagcgct agtgattacg ccgcagattc ccatggtaac agaagagttt
     1021 gtgaaaccat cacagggcca tgtgaccttt gaggatattg ctgtgtactt ctcccaggag
     1081 gagtggggcc tccttgatga agctcaaagg tgcctgtatc atgatgtgat gctggagaac
     1141 ttttcgctta tggcctcagt aggttgtttg catggaatag aggctgagga ggccccttct
     1201 gagcagactc tttctgcgca aggagtgtca caggccagga ctccaaagct aggtccttcc
     1261 atcccaaatg ctcattcttg tgagatgtgt atcctggtca tgaaagacat tttgtacctc
     1321 agtgagcatc aggggacact tccctggcag aaaccttata cgtctgtggc cagtgggaaa
     1381 tggttttcat ttggttctaa cctgcaacag caccagaacc aggacagtgg agagaaacac
     1441 atcagaaagg aggagagcag tgccttgctt ctgaatagct gcaaaattcc tctgtcagac
     1501 aatcttttcc catgcaaaga tgttgagaag gattttccaa ccatcctggg ccttctccaa
     1561 caccagacca cccacagcag acaagagtat gcacatagaa gcagggagac ctttcaacaa
     1621 agacgttaca aatgtgagca agttttcaat gagaaagttc atgttactga gcatcagaga
     1681 gtccacactg gagaaaaagc ttataagcgt agggaatatg ggaaatcctt gaactctaaa
     1741 tacttatttg ttgaacacca gagaacccat aatgcagaaa agccttatgt gtgcaatata
     1801 tgtgggaaat cattcctcca taaacaaaca ctcgttgggc accagcagag aattcacact
     1861 agagaaaggt cttatgtgtg catcgaatgt gggaaatcct tgagctccaa atactcactt
     1921 gtggaacacc agagaaccca taatggagaa aagccttatg tgtgcaatgt atgtgggaaa
     1981 tcattccgcc acaaacaaac atttgttggc catcagcaga gaatccacac tggagagagg
     2041 ccttatgtgt gtatggaatg tgggaaatct tttattcatt cctatgaccg cattcgacac
     2101 cagagagttc acactggaga aggggcttat cagtgcagtg aatgtgggaa atccttcata
     2161 tacaaacagt cacttcttga tcaccataga atccacacgg gagaaaggcc ttatgagtgc
     2221 aaagaatgtg ggaaggcctt cattcacaaa aaaagacttc ttgagcacca gagaattcat
     2281 actggagaaa agccttatgt gtgcatcata tgtgggaaat catttatccg ctcgtctgac
     2341 tacatgcgac accagagaat tcacactgga gaaagggctt atgaatgcag tgactgtggg
     2401 aaagccttca tctccaaaca aacacttctt aagcatcaca aaatccacac tagagaaagg
     2461 ccttatgaat gcagtgaatg tggaaaaggc ttctaccttg aggttaaact tcttcagcac
     2521 caaagaatcc atactagaga acaactttgt gagtgcaatg aatgtggaaa agtcttcagc
     2581 caccaaaaaa gacttcttga gcaccagaaa gttcacactg gcgaaaagcc ctgtgagtgc
     2641 agtgaatgtg ggaaatgctt tagacaccgc accagcctca ttcaacacca gaaagttcac
     2701 agtggagaga ggccttataa ctgcactgca tgtgagaagg cctttatcta taaaaacaaa
     2761 cttgttgagc atcagcgaat ccacaccgga gaaaagccgt atgaatgtgg taaatgtggg
     2821 aaagccttca acaaaagata ttcccttgtc aggcaccaga aggtacatat aacagaagag
     2881 cccctcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     2941 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3001 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3061 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3121 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3181 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3241 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3301 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3361 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3421 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3481 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3541 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3601 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3661 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3721 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3781 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3841 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3901 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3961 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4021 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4081 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4141 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4201 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4261 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4321 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4381 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4441 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4501 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4561 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4621 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4681 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4741 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4801 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4861 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4921 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4981 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5041 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5101 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5161 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5221 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5281 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5341 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5401 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5461 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5521 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5581 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5641 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5701 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5761 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5821 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5881 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5941 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6001 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6061 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6121 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6241 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6361 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6421 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6481 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6541 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6601 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6661 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6721 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6781 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6841 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6901 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6961 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7021 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7081 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7141 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7201 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7261 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7321 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7381 gccacctgac gtc
//
