LOCUS       V035332                 8035 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035332
VERSION     V035332
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      880..889
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             889..954
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             958..3717
                     /label="AR(NM_000044)"
                     /note="AR(NM_000044)"
                     /gene="AR"
     polyA_signal    3797..4021
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4067..4495
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4509..4838
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4905..5504
                     /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    5678..5799
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5848..5864)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5872..5888
                     /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(5896..5926)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5941..5962
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6250..6835)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7006..7866)
                     /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(7867..7971)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag ccaccatgga ctacaaagac
      901 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagcttatg
      961 gaagtgcagt tagggctggg aagggtctac cctcggccgc cgtccaagac ctaccgagga
     1021 gctttccaga atctgttcca gagcgtgcgc gaagtgatcc agaacccggg ccccaggcac
     1081 ccagaggccg cgagcgcagc acctcccggc gccagtttgc tgctgctgca gcagcagcag
     1141 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcaagag
     1201 actagcccca ggcagcagca gcagcagcag ggtgaggatg gttctcccca agcccatcgt
     1261 agaggcccca caggctacct ggtcctggat gaggaacagc aaccttcaca gccgcagtcg
     1321 gccctggagt gccaccccga gagaggttgc gtcccagagc ctggagccgc cgtggccgcc
     1381 agcaaggggc tgccgcagca gctgccagca cctccggacg aggatgactc agctgcccca
     1441 tccacgttgt ccctgctggg ccccactttc cccggcttaa gcagctgctc cgctgacctt
     1501 aaagacatcc tgagcgaggc cagcaccatg caactccttc agcaacagca gcaggaagca
     1561 gtatccgaag gcagcagcag cgggagagcg agggaggcct cgggggctcc cacttcctcc
     1621 aaggacaatt acttaggggg cacttcgacc atttctgaca acgccaagga gttgtgtaag
     1681 gcagtgtcgg tgtccatggg cctgggtgtg gaggcgttgg agcatctgag tccaggggaa
     1741 cagcttcggg gggattgcat gtacgcccca cttttgggag ttccacccgc tgtgcgtccc
     1801 actccttgtg ccccattggc cgaatgcaaa ggttctctgc tagacgacag cgcaggcaag
     1861 agcactgaag atactgctga gtattcccct ttcaagggag gttacaccaa agggctagaa
     1921 ggcgagagcc taggctgctc tggcagcgct gcagcaggga gctccgggac acttgaactg
     1981 ccgtctaccc tgtctctcta caagtccgga gcactggacg aggcagctgc gtaccagagt
     2041 cgcgactact acaactttcc actggctctg gccggaccgc cgccccctcc gccgcctccc
     2101 catccccacg ctcgcatcaa gctggagaac ccgctggact acggcagcgc ctgggcggct
     2161 gcggcggcgc agtgccgcta tggggacctg gcgagcctgc atggcgcggg tgcagcggga
     2221 cccggttctg ggtcaccctc agccgccgct tcctcatcct ggcacactct cttcacagcc
     2281 gaagaaggcc agttgtatgg accgtgtggt ggtggtgggg gtggtggcgg cggcggcggc
     2341 ggcggcggcg gcggcggcgg cggcggcggc ggcggcgagg cgggagctgt agccccctac
     2401 ggctacactc ggccccctca ggggctggcg ggccaggaaa gcgacttcac cgcacctgat
     2461 gtgtggtacc ctggcggcat ggtgagcaga gtgccctatc ccagtcccac ttgtgtcaaa
     2521 agcgaaatgg gcccctggat ggatagctac tccggacctt acggggacat gcgtttggag
     2581 actgccaggg accatgtttt gcccattgac tattactttc caccccagaa gacctgcctg
     2641 atctgtggag atgaagcttc tgggtgtcac tatggagctc tcacatgtgg aagctgcaag
     2701 gtcttcttca aaagagccgc tgaagggaaa cagaagtacc tgtgcgccag cagaaatgat
     2761 tgcactattg ataaattccg aaggaaaaat tgtccatctt gtcgtcttcg gaaatgttat
     2821 gaagcaggga tgactctggg agcccggaag ctgaagaaac ttggtaatct gaaactacag
     2881 gaggaaggag aggcttccag caccaccagc cccactgagg agacaaccca gaagctgaca
     2941 gtgtcacaca ttgaaggcta tgaatgtcag cccatctttc tgaatgtcct ggaagccatt
     3001 gagccaggtg tagtgtgtgc tggacacgac aacaaccagc ccgactcctt tgcagccttg
     3061 ctctctagcc tcaatgaact gggagagaga cagcttgtac acgtggtcaa gtgggccaag
     3121 gccttgcctg gcttccgcaa cttacacgtg gacgaccaga tggctgtcat tcagtactcc
     3181 tggatggggc tcatggtgtt tgccatgggc tggcgatcct tcaccaatgt caactccagg
     3241 atgctctact tcgcccctga tctggttttc aatgagtacc gcatgcacaa gtcccggatg
     3301 tacagccagt gtgtccgaat gaggcacctc tctcaagagt ttggatggct ccaaatcacc
     3361 ccccaggaat tcctgtgcat gaaagcactg ctactcttca gcattattcc agtggatggg
     3421 ctgaaaaatc aaaaattctt tgatgaactt cgaatgaact acatcaagga actcgatcgt
     3481 atcattgcat gcaaaagaaa aaatcccaca tcctgctcaa gacgcttcta ccagctcacc
     3541 aagctcctgg actccgtgca gcctattgcg agagagctgc atcagttcac ttttgacctg
     3601 ctaatcaagt cacacatggt gagcgtggac tttccggaaa tgatggcaga gatcatctct
     3661 gtgcaagtgc ccaagatcct ttctgggaaa gtcaagccca tctatttcca cacccagtga
     3721 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3781 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3841 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3901 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3961 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4021 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4081 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4141 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4201 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4261 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4321 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4381 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4441 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4501 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4561 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4621 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4681 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4741 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4801 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4861 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgacc gagtacaagc
     4921 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     4981 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     5041 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     5101 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     5161 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     5221 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     5281 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     5341 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     5401 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     5461 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgagcggga ctctggggtt
     5521 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5581 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5641 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5701 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5761 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5821 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5881 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5941 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6001 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6061 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6661 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6781 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7141 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7861 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7981 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
