LOCUS       V025891                 7933 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025891
VERSION     V025891
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..3684
                     /label="AR(NM_000044)"
                     /note="AR(NM_000044)"
                     /gene="AR"
     CDS             3697..3762
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     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             4850..5449
                     /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    5609..5730
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5779..5795)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5803..5819
                     /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(5827..5857)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5872..5893
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6181..6766)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6937..7797)
                     /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(7798..7902)
                     /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 caccatggaa gtgcagttag ggctgggaag ggtctaccct
      961 cggccgccgt ccaagaccta ccgaggagct ttccagaatc tgttccagag cgtgcgcgaa
     1021 gtgatccaga acccgggccc caggcaccca gaggccgcga gcgcagcacc tcccggcgcc
     1081 agtttgctgc tgctgcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     1141 cagcagcagc agcagcagca gcaagagact agccccaggc agcagcagca gcagcagggt
     1201 gaggatggtt ctccccaagc ccatcgtaga ggccccacag gctacctggt cctggatgag
     1261 gaacagcaac cttcacagcc gcagtcggcc ctggagtgcc accccgagag aggttgcgtc
     1321 ccagagcctg gagccgccgt ggccgccagc aaggggctgc cgcagcagct gccagcacct
     1381 ccggacgagg atgactcagc tgccccatcc acgttgtccc tgctgggccc cactttcccc
     1441 ggcttaagca gctgctccgc tgaccttaaa gacatcctga gcgaggccag caccatgcaa
     1501 ctccttcagc aacagcagca ggaagcagta tccgaaggca gcagcagcgg gagagcgagg
     1561 gaggcctcgg gggctcccac ttcctccaag gacaattact tagggggcac ttcgaccatt
     1621 tctgacaacg ccaaggagtt gtgtaaggca gtgtcggtgt ccatgggcct gggtgtggag
     1681 gcgttggagc atctgagtcc aggggaacag cttcgggggg attgcatgta cgccccactt
     1741 ttgggagttc cacccgctgt gcgtcccact ccttgtgccc cattggccga atgcaaaggt
     1801 tctctgctag acgacagcgc aggcaagagc actgaagata ctgctgagta ttcccctttc
     1861 aagggaggtt acaccaaagg gctagaaggc gagagcctag gctgctctgg cagcgctgca
     1921 gcagggagct ccgggacact tgaactgccg tctaccctgt ctctctacaa gtccggagca
     1981 ctggacgagg cagctgcgta ccagagtcgc gactactaca actttccact ggctctggcc
     2041 ggaccgccgc cccctccgcc gcctccccat ccccacgctc gcatcaagct ggagaacccg
     2101 ctggactacg gcagcgcctg ggcggctgcg gcggcgcagt gccgctatgg ggacctggcg
     2161 agcctgcatg gcgcgggtgc agcgggaccc ggttctgggt caccctcagc cgccgcttcc
     2221 tcatcctggc acactctctt cacagccgaa gaaggccagt tgtatggacc gtgtggtggt
     2281 ggtgggggtg gtggcggcgg cggcggcggc ggcggcggcg gcggcggcgg cggcggcggc
     2341 ggcgaggcgg gagctgtagc cccctacggc tacactcggc cccctcaggg gctggcgggc
     2401 caggaaagcg acttcaccgc acctgatgtg tggtaccctg gcggcatggt gagcagagtg
     2461 ccctatccca gtcccacttg tgtcaaaagc gaaatgggcc cctggatgga tagctactcc
     2521 ggaccttacg gggacatgcg tttggagact gccagggacc atgttttgcc cattgactat
     2581 tactttccac cccagaagac ctgcctgatc tgtggagatg aagcttctgg gtgtcactat
     2641 ggagctctca catgtggaag ctgcaaggtc ttcttcaaaa gagccgctga agggaaacag
     2701 aagtacctgt gcgccagcag aaatgattgc actattgata aattccgaag gaaaaattgt
     2761 ccatcttgtc gtcttcggaa atgttatgaa gcagggatga ctctgggagc ccggaagctg
     2821 aagaaacttg gtaatctgaa actacaggag gaaggagagg cttccagcac caccagcccc
     2881 actgaggaga caacccagaa gctgacagtg tcacacattg aaggctatga atgtcagccc
     2941 atctttctga atgtcctgga agccattgag ccaggtgtag tgtgtgctgg acacgacaac
     3001 aaccagcccg actcctttgc agccttgctc tctagcctca atgaactggg agagagacag
     3061 cttgtacacg tggtcaagtg ggccaaggcc ttgcctggct tccgcaactt acacgtggac
     3121 gaccagatgg ctgtcattca gtactcctgg atggggctca tggtgtttgc catgggctgg
     3181 cgatccttca ccaatgtcaa ctccaggatg ctctacttcg cccctgatct ggttttcaat
     3241 gagtaccgca tgcacaagtc ccggatgtac agccagtgtg tccgaatgag gcacctctct
     3301 caagagtttg gatggctcca aatcaccccc caggaattcc tgtgcatgaa agcactgcta
     3361 ctcttcagca ttattccagt ggatgggctg aaaaatcaaa aattctttga tgaacttcga
     3421 atgaactaca tcaaggaact cgatcgtatc attgcatgca aaagaaaaaa tcccacatcc
     3481 tgctcaagac gcttctacca gctcaccaag ctcctggact ccgtgcagcc tattgcgaga
     3541 gagctgcatc agttcacttt tgacctgcta atcaagtcac acatggtgag cgtggacttt
     3601 ccggaaatga tggcagagat catctctgtg caagtgccca agatcctttc tgggaaagtc
     3661 aagcccatct atttccacac ccagctcgag accggtgact acaaagacca tgacggtgat
     3721 tataaagatc atgacatcga ttacaaggat gacgatgaca agtgaccggt aagtttaaac
     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 gattttggcc
     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 tgaccgagta
     4861 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4921 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4981 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5041 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5101 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5161 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5221 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5281 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     5341 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5401 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat tcgaaatgac
     5461 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5521 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5581 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5701 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5761 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5821 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5881 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5941 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6001 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6061 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6121 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6181 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6301 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6361 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6421 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6481 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6541 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6601 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6661 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6721 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6781 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6841 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6901 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6961 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7021 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7081 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7141 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7201 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7261 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7321 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7381 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7441 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7501 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7561 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7621 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7681 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7741 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7801 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7861 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7921 gccacctgac gtc
//
