LOCUS       V032221                 6751 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032221
VERSION     V032221
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             925..2703
                     /label="Cdyl(NM_009881)"
                     /note="Cdyl(NM_009881)"
                     /gene="Cdyl"
     CDS             2716..2781
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2816..3040
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3086..3514
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3528..3857
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3869..4267
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     polyA_signal    4427..4548
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4597..4613)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4621..4637
                     /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(4645..4675)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4690..4711
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4999..5584)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5755..6615)
                     /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(6616..6720)
                     /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 gagaattcgc caccatgggc ataggcaata gccagcctaa ttcacaggaa
      961 gcccagctct gcacacttcc agagaaagct gaacaaccta ctgatgataa cacctgccag
     1021 caaaataatg tggttcctgc aacagtctca gaacccgatc aagcgtcccc tgcaattcaa
     1081 gacgcggaga ctcaggtgga aagtatcgtt gacaaaagga aaaacaagaa agggaagaca
     1141 gaatatctgg tgcggtggaa aggctatgac agtgaggatg acacgtggga gcctgagcag
     1201 cacctggtga actgtgagga atacatccat gacttcaacc ggcgccacaa cgagaggcaa
     1261 aaggaaggta gcctggctcg tgccagcaga gcctccccca gcaacgcccg gaagcagatt
     1321 tccaggtcca cccacagcac tctctccaag accaactcca aagcacttgt ggtaggcaaa
     1381 gatcatgagt ccaaaagcag ccagctgttg gctgccagcc agaagttcag gaaaaaccca
     1441 gccccatctc ttgcaaaccg caagaacatg gacctcgcca agtcagggat caaaattctc
     1501 gtgcctaaga gccccgttaa gggcaggacc tcggttgatg gctttcaggg ggagagcccc
     1561 gagaagctgg accctgtgga tcagggtgcc gaggacactg tagccccaga ggtgactgca
     1621 gagaagccca ctggggcttt gctgggccct ggtgcggagc gagccaggat ggggagcagg
     1681 ccccgaatac atccactagt gcctcaggtt tctggccccg tgactgctgc catggccaca
     1741 ggcttagctg ttaatggaaa aggtacatct ccattcatgg atgcgctagc agccaacgga
     1801 acagtcacca tacagacatc cgtaacagga gtgacagccg ggaaaaggaa atttattgac
     1861 gacagaagag accaaccttt tgacaagcgg ttgcgtttca gtgtgaggca gacagagagt
     1921 gcctacagat acagagatat tgtcgtcagg aagcaagatg gcttcaccca catcttgtta
     1981 tccacaaaat cgtcagagaa taactcacta aacccagagg tgatgaaaga agtgcagagc
     2041 gccctgagca cagctgcagc cgacgacagc aagctggttc tgctcagcgc cgtgggcagc
     2101 gtcttctgct gtggtctgga ctttatttat tttattcggc gcctcacaga tgaccgaaag
     2161 agagaaagca ctaaaatggc agacgctatc agaaacttcg tgaatacttt cattcagttt
     2221 aagaagccta ttattgtagc tgttaatggc ccagccattg gactaggagc atccatattg
     2281 cctctttgtg atgtggtttg ggctaacgaa aaggcttggt ttcaaacacc ctataccacc
     2341 ttcggacaga gtccagatgg ctgctctacc gttatgtttc ccaagattat gggaggagca
     2401 tctgcgaatg aaatgctgtt cagtgggcgg aagttgacgg cacaggaggc ctgtggcaag
     2461 ggtctggtct cccaggtgtt ttggccagga accttcacac aggaagtcat ggttcgaatc
     2521 aaggagctgg cttcatgtaa cccagttgtc ctggaggaat ccaaagccct ggtgcgctgc
     2581 aatatgaaga tggagctaga gcaggccaat gagagagaat gtgaagtgct gaagaagatc
     2641 tggggctccg cccagggcat ggactccatg ttaaagtact tacagaggaa aatcgatgag
     2701 ttcctcgaga ccggtgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     2761 tacaaggatg acgatgacaa gtgaccggta agtttaaacc gctgatcagc ctcgactgtg
     2821 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2881 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2941 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3001 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3061 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3241 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3301 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3361 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3421 atctcggtct attcttttga tttataaggg attttggcca tttcggccta ttggttaaaa
     3481 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3601 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3661 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3721 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3781 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3841 gcctaggctt ttgcaaaaag ctcccgggat ggccaagcct ttgtctcaag aagaatccac
     3901 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     3961 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     4021 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     4081 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     4141 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     4201 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     4261 gggctaattc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4561 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5401 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5521 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5641 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5701 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5761 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5821 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5881 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5941 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6001 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6061 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6121 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6181 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6241 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6301 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6361 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6421 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6481 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6541 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6601 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6661 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6721 cacatttccc cgaaaagtgc cacctgacgt c
//
