LOCUS       V033684                 7384 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033684
VERSION     V033684
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             919..3135
                     /label="SUZ12(NM_015355)"
                     /note="SUZ12(NM_015355)"
                     /gene="SUZ12"
     CDS             3148..3213
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3248..3472
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3518..3946
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3960..4289
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4301..4900
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5060..5181
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5230..5246)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5254..5270
                     /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(5278..5308)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5323..5344
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5632..6217)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6388..7248)
                     /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(7249..7353)
                     /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 aagctttcta gagccaccat ggcgcctcag aagcacggcg gtgggggagg gggcggctcg
      961 gggcccagcg cggggtccgg gggaggcggc ttcgggggtt cggcggcggt ggcggcggcg
     1021 acggcttcgg gcggcaaatc cggcggcggg agctgtggag ggggtggcag ttactcggcc
     1081 tcctcctcct cctccgcggc ggcagcggcg ggggctgcgg tgttaccggt gaagaagccg
     1141 aaaatggagc acgtccaggc tgaccacgag cttttcctcc aggcctttga gaagccaaca
     1201 cagatctata gatttcttcg aactcggaat ctcatagcac caatattttt gcacagaact
     1261 cttacttaca tgtctcatcg aaactccaga acaaacatca aaaggaaaac atttaaagtt
     1321 gatgatatgt tatcaaaagt agagaaaatg aaaggagagc aagaatctca tagcttgtca
     1381 gctcatttgc agcttacgtt tactggtttc ttccacaaaa atgataagcc atcaccaaac
     1441 tcagaaaatg aacaaaattc tgttaccctg gaagtcctgc ttgtgaaagt ttgccacaaa
     1501 aaaagaaagg atgtaagttg tccaataagg caagttccca caggtaaaaa gcaggtgcct
     1561 ttgaatcctg acctcaatca aacaaaaccc ggaaatttcc cgtcccttgc agtttccagt
     1621 aatgaatttg aacctagtaa cagccatatg gtgaagtctt actcgttgct atttagagtg
     1681 actcgtccag gaagaagaga gtttaatgga atgattaatg gagaaaccaa tgaaaatatt
     1741 gatgtcaatg aagagcttcc agccagaaga aaacgaaatc gtgaggatgg ggaaaagaca
     1801 tttgttgcac aaatgacagt atttgataaa aacaggcgct tacagctttt agatggggaa
     1861 tatgaagtag ccatgcagga aatggaagaa tgtccaataa gcaagaaaag agcaacatgg
     1921 gagactattc ttgatgggaa gaggctgcct ccattcgaaa cattttctca gggacctacg
     1981 ttgcagttca ctcttcgttg gacaggagag accaatgata aatctacggc tcctattgcc
     2041 aaacctcttg ccactagaaa ttcagagagt ctccatcagg aaaacaagcc tggttcagtt
     2101 aaacctactc aaactattgc tgttaaagaa tcattgacta cagatctaca aacaagaaaa
     2161 gaaaaggata ctccaaatga aaaccgacaa aaattaagaa tattttatca gtttctctat
     2221 aacaacaata caaggcaaca aactgaagca agagatgacc tgcattgccc ttggtgtact
     2281 ctgaactgcc gcaaacttta tagtttactc aagcatctta aactctgcca tagcagattt
     2341 atcttcaact atgtttatca tccaaaaggt gctaggatag atgtttctat caatgagtgt
     2401 tatgatggct cctatgcagg aaatcctcag gatattcatc gccaacctgg atttgctttt
     2461 agtcgcaacg gaccagttaa gagaacacct atcacacata ttcttgtgtg caggccaaaa
     2521 cgaacaaaag caagcatgtc tgaatttctt gaatctgaag atggggaagt agaacagcaa
     2581 agaacatata gtagtggcca caatcgtctg tatttccata gtgatacctg cttacctctc
     2641 cgtccacaag aaatggaagt agatagtgaa gatgaaaagg atcctgaatg gctaagagaa
     2701 aaaaccatta cacaaattga agagttttct gatgttaatg aaggagagaa agaagtgatg
     2761 aaactctgga atctccatgt catgaagcat gggtttattg ctgacaatca aatgaatcat
     2821 gcctgtatgc tgtttgtaga aaattatgga cagaaaataa ttaagaagaa tttatgtcga
     2881 aacttcatgc ttcatctagt cagcatgcat gactttaatc ttattagcat aatgtcaata
     2941 gataaagctg ttaccaagct ccgtgaaatg cagcaaaaat tagaaaaggg ggaatctgct
     3001 tcccctgcaa acgaagaaat aactgaagaa caaaatggga cagcaaatgg atttagtgaa
     3061 attaactcaa aagagaaagc tttggaaaca gatagtgtct caggggtttc aaaacagagc
     3121 aaaaaacaaa aactcctcga gaccggtgac tacaaagacc atgacggtga ttataaagat
     3181 catgacatcg attacaagga tgacgatgac aagtgaccgg taagtttaaa ccgctgatca
     3241 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3301 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3361 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3421 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3481 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggc catttcggcc
     3901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4201 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4261 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg atgaccgagt acaagcccac
     4321 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4381 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4441 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4501 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4561 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4621 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4681 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4741 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4801 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4861 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc
     4921 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4981 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5041 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5101 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5161 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5221 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5281 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5341 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5401 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5461 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5521 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5581 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5641 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5701 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5761 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5821 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5881 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5941 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6001 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6061 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6121 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7381 cgtc
//
