LOCUS       V033149                 7075 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033149
VERSION     V033149
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             937..2826
                     /label="MAN1C1(NM_020379)"
                     /note="MAN1C1(NM_020379)"
                     /gene="MAN1C1"
     CDS             2839..2904
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2939..3163
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3209..3637
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3651..3980
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3992..4591
                     /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    4751..4872
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4921..4937)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4945..4961
                     /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(4969..4999)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5014..5035
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5323..5908)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6079..6939)
                     /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(6940..7044)
                     /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 gagaattcga tatcggatcc gccaccatgc tcatgaggaa agtgcccggc
      961 ttcgtcccgg cctccccgtg ggggctgcgg ctgccgcaga agttcctctt cctcctcttc
     1021 ctctcgggcc tggtcaccct gtgcttcggg gccctcttcc tgctgcccca ctcctctcgc
     1081 ctcaagcgcc tcttcctggc cccccggacc cagcagcctg gtctggaagt ggtggctgaa
     1141 atcgccggcc atgccccggc ccgcgagcag gagccgcctc ccaacccggc ccccgccgcg
     1201 ccggccccgg gcgaggatga ccccagcagc tgggccagtc cccgccgcag gaaagggggg
     1261 ctgcggcgca cccgccccac tggaccccgc gaggaggcca cggcggcccg gggcaatagc
     1321 atcccggcct ccaggcccgg ggacgagggc gtccctttcc gctttgactt caacgcattc
     1381 cggagccgtc tccgccaccc ggtcctggga acgagggccg atgagagtca ggagccccag
     1441 agccaagtgc gagcccagcg ggagaaaatc aaggagatga tgcagtttgc ttggcagagc
     1501 tataagcgtt atgcaatggg gaaaaacgaa ctccgtccac taacaaaaga tggctacgag
     1561 ggtaacatgt tcggaggcct cagcggggca acagtcattg actccctcga taccctctac
     1621 ctcatggagc tgaaggagga gttccaggag gccaaggcct gggtgggaga gagcttccac
     1681 ctgaacgtga gcggagaagc atccttgttt gaggtgaaca tccgctacat cgggggactc
     1741 ctctcagcct tctacctgac aggagaagag gtgttccgaa taaaggccat caggctggga
     1801 gagaagctcc tgccggcgtt caacaccccc acgggaatcc caaagggcgt ggtgagcttc
     1861 aaaagtggga actggggctg ggccacagcc ggcagcagca gcatcttggc ggagtttgga
     1921 tccctgcact tggaattctt acacctcact gaactctctg gcaaccaggt cttcgctgaa
     1981 aaggtcagga acatccgcaa ggtcctcagg aagatcgaaa agccctttgg cctctacccc
     2041 aacttcctca gcccagtgag tgggaactgg gtgcaacacc atgtctcagt tggaggactc
     2101 ggggacagtt tttatgaata tttgatcaaa tcctggttga tgtcgggcaa gacagatatg
     2161 gaggctaaaa atatgtacta cgaagccttg gaggcgatag agacctactt gctgaatgtc
     2221 tctcccgggg ggctgaccta cattgccgag tggcgagggg ggattctgga ccacaagatg
     2281 gggcacctgg cctgtttctc cgggggcatg atcgcccttg gcgccgagga tgccaaggaa
     2341 gaaaagaggg cccactaccg agagctcgca gcccagatca ccaagacgtg tcacgagtca
     2401 tacgcccgct cagacaccaa acttgggcct gaggccttct ggtttaactc cggcagagag
     2461 gccgtggcca cccagctgag cgagagctac tacatcctcc ggccagaggt ggtggagagc
     2521 tacatgtacc tgtggcgaca gacccacaac cccatctaca gggagtgggg ctgggaggtg
     2581 gtgctggcct tggagaaata ctgtcggaca gaagccggtt tctctgggat ccaagacgtg
     2641 tacagtagca cccccaacca cgacaacaag cagcagagct tctttctagc ggagacacta
     2701 aagtatctct atcttctgtt ctctgaagat gacttgctct ccctggaaga ctgggtgttc
     2761 aacaccgagg cccacccact cccggtgaac cactcagaca gctccggcag agcctggggc
     2821 agacacctcg agaccggtga ctacaaagac catgacggtg attataaaga tcatgacatc
     2881 gattacaagg atgacgatga caagtgaccg gtaagtttaa accgctgatc agcctcgact
     2941 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3001 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3061 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3121 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3181 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3241 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3301 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3361 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3421 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3481 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3541 cctatctcgg tctattcttt tgatttataa gggattttgg ccatttcggc ctattggtta
     3601 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3661 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3721 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3781 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3841 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3901 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3961 gaggcctagg cttttgcaaa aagctcccgg gatgaccgag tacaagccca cggtgcgcct
     4021 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     4081 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     4141 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4201 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4261 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4321 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4381 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4441 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4501 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4561 ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg accgaccaag cgacgcccaa
     4621 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4681 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4741 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4801 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4861 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4921 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4981 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5041 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5101 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5161 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5221 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5281 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5341 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5521 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5581 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5641 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5701 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5761 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5821 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     5881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6121 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6181 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6241 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6301 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6361 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6421 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6481 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6541 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6601 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6661 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6721 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6781 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6841 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6901 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6961 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7021 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
