LOCUS       V044523                 5515 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044523
VERSION     V044523
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..1308
                     /label="Rpl22(NM_009079)"
                     /note="Rpl22(NM_009079)"
                     /gene="Rpl22"
     CDS             1309..1335
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    1379..1603
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      1649..2077
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2091..2420
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2432..3031
                     /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    3191..3312
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3361..3377)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3385..3401
                     /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(3409..3439)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3454..3475
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3763..4348)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4519..5379)
                     /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(5380..5484)
                     /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 caccatggcg cctgtgaaaa agcttgtggc gaaggggggc
      961 aaaaaaaaga agcaggtttt gaagttcacc ctggactgca ctcaccctgt agaagatgga
     1021 atcatggatg ctgccaattt tgagcagttc ctccaggaga gaatcaaggt gaatgggaaa
     1081 gctggcaacc tcggcggagg agtcgtgacc atcgaacgca gcaagagcaa gatcactgtc
     1141 acttcagagg tgcctttctc caaaaggtat ttgaaatatc tcaccaaaaa atatttgaag
     1201 aagaacaacc tccgagactg gctgcgtgtt gtcgccaaca gcaaagagag ttacgagctg
     1261 cgttacttcc agattaacca ggatgaagag gaggaggaag acgaggatta cccatacgat
     1321 gttccagatt acgcttagct cgagtgaccg gtaagtttaa accgctgatc agcctcgact
     1381 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1441 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1501 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     1561 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     1621 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     1681 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1741 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1801 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1861 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1921 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1981 cctatctcgg tctattcttt tgatttataa gggattttgg ccatttcggc ctattggtta
     2041 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2101 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2161 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2221 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2281 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2341 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2401 gaggcctagg cttttgcaaa aagctcccgg gatgaccgag tacaagccca cggtgcgcct
     2461 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     2521 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     2581 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     2641 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     2701 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     2761 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     2821 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     2881 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     2941 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     3001 ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg accgaccaag cgacgcccaa
     3061 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3121 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3181 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3241 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3301 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     3361 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3421 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4201 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     4321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4621 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
