LOCUS       V041493                 6798 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041493
VERSION     V041493
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..2457
                     /label="ABAT(NM_020686)"
                     /note="ABAT(NM_020686)"
                     /gene="ABAT"
     polyA_signal    2560..2784
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2830..3258
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3272..3601
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3668..4267
                     /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    4441..4562
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4611..4627)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4635..4651
                     /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(4659..4689)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4704..4725
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5013..5598)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5769..6629)
                     /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(6630..6734)
                     /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 gcctccatgt tgctcgccca gcgcctggcc tgcagcttcc agcacagcta ccgcctgctg
     1021 gtgcctggat ccagacacat tagtcaagct gcagccaaag tcgacgttga atttgattat
     1081 gatgggcctc tgatgaagac ggaagtccca gggcctagat ctcaggagtt aatgaaacag
     1141 ctgaatataa ttcagaatgc agaggctgtg cattttttct gcaattacga agagagccga
     1201 ggcaattacc tggttgatgt ggacggcaac cgaatgctgg atctttattc ccagatctcc
     1261 tctgtcccca taggttacag ccaccccgcc ctgctgaaac tcatccaaca gcctcaaaat
     1321 gcgagcatgt ttgtcaacag acccgccctc ggaatcctgc ctccggagaa ctttgtggag
     1381 aagctccggc agtccttgct ctcggtggct cccaaaggga tgtcccagct catcaccatg
     1441 gcctgcggct cctgctccaa tgaaaacgcc ttaaagacca tcttcatgtg gtaccggagc
     1501 aaggaaagag ggcagagggg cttctcccag gaggagctgg agacgtgcat gattaaccag
     1561 gcccctggct gccccgacta cagcatcctc tccttcatgg gcgcgttcca tgggaggacc
     1621 atgggttgct tagcgaccac gcactctaaa gccattcaca agatcgacat cccttccttt
     1681 gactggccca tcgcaccgtt cccacggctg aaataccctc tggaagagtt tgtgaaagag
     1741 aaccaacagg aggaggcccg ctgtctggaa gaggtggagg atctgattgt gaaatatcgg
     1801 aaaaagaaga agacggtggc cgggatcatc gtggagccca tccagtccga gggtggagac
     1861 aaccacgcat ccgatgactt ctttcggaag ctgagagaca tcgccaggaa gcatggctgc
     1921 gccttcttgg tggacgaggt ccagaccgga ggaggctgca cgggcaagtt ctgggcccat
     1981 gagcactggg gcctggatga cccagcagac gtgatgacct tcagcaagaa gatgatgact
     2041 gggggcttct tccacaagga ggagttcagg cctaatgctc cctaccggat cttcaacacc
     2101 tggctggggg acccgtccaa gaacctgttg ctggctgagg tcatcaacat catcaagcgg
     2161 gaggacctgc taaataatgc agcccatgcc gggaaggccc tgctcacagg actgctggac
     2221 ctccaggccc ggtaccccca gttcatcagc agggtgagag gacgaggcac cttttgctcc
     2281 ttcgatactc ccgatgattc catacggaat aagctcattt taattgccag aaacaaaggt
     2341 gtggtgttgg gtggctgtgg tgacaaatcc attcgtttcc gtcccacgct ggtcttcagg
     2401 gatcaccacg ctcacctgtt cctcaatatt ttcagtgaca tcttagcaga cttcaagtaa
     2461 ggatccacta gtccagtgtg gtggaattct gcagatatcc agcacagtgg cggccgctcg
     2521 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     2581 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2641 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2701 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2761 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2821 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2881 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2941 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3001 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3061 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3121 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3181 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3241 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3361 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3421 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3481 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3541 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3601 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3661 gtttcgcatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     3721 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3781 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     3841 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     3901 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     3961 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4021 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4081 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4141 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4201 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4261 tgcctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4321 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4381 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4441 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4501 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4561 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5521 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     5581 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5641 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5701 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5761 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5821 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5881 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5941 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6001 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6061 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6121 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6181 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6241 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6301 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6361 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6421 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6481 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6541 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6601 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6661 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6721 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     6781 ctcccgatcc cctatggt
//
