LOCUS       V028629                 6667 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028629
VERSION     V028629
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..2418
                     /label="USP14(NM_005151)"
                     /note="USP14(NM_005151)"
                     /gene="USP14"
     CDS             2431..2496
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2531..2755
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2801..3229
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3243..3572
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3584..4183
                     /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    4343..4464
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4513..4529)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4537..4553
                     /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(4561..4591)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4606..4627
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4915..5500)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5671..6531)
                     /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(6532..6636)
                     /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 cgctctactc cgttactgta
      961 aaatggggaa aggagaaatt tgaaggtgta gaattgaata cagatgaacc tccaatggta
     1021 ttcaaggctc agctgtttgc gttgactgga gtccagcctg ccagacagaa agttatggtg
     1081 aaaggaggaa cgctaaagga tgatgattgg ggaaacatca aaataaaaaa tggaatgact
     1141 ctactaatga tggggtcagc agatgctctt ccagaagaac cctcagccaa aactgtcttc
     1201 gtagaagaca tgacagaaga acagttagca tctgctatgg agttaccatg tggattgaca
     1261 aaccttggta acacttgtta catgaatgcc acagttcagt gtattcgttc tgtgcctgaa
     1321 ctcaaagatg cccttaaaag gtatgcaggt gccttgagag cttcagggga aatggcttca
     1381 gcgcagtata ttactgcagc ccttagagat ttgtttgatt ccatggataa aacttcttcc
     1441 agtattccac ctattattct actgcagttt ttgcacatgg ctttcccaca gtttgccgag
     1501 aaaggtgaac aaggacagta tcttcaacag gatgctaatg aatgttggat acaaatgatg
     1561 cgagtattgc aacagaaatt ggaagcaata gaggatgatt ctgttaaaga gacagactcc
     1621 tcatctgcat cggcagcgac accttctaaa aagaaaagtt taatcgatca gttcttcggt
     1681 gttgagtttg aaactaccat gaaatgtaca gaatctgaag aagaagaagt caccaaagga
     1741 aaggaaaatc aacttcagct tagctgtttt atcaatcagg aagtcaagta tctttttaca
     1801 ggacttaaat tgcgacttca ggaagaaatc accaaacagt ctccaacgtt gcaaagaaat
     1861 gccttgtata tcaaatcttc caagatcagc cggctgcctg cttacttgac cattcagatg
     1921 gttcgatttt tttataaaga gaaggaatct gtgaatgcca aagttcttaa ggatgttaaa
     1981 tttcctctta tgttggatat gtatgaactg tgtacaccag aacttcaaga gaaaatggtg
     2041 tcttttcgat ccaaattcaa ggatctagaa gataaaaaag tgaatcagca gccaaataca
     2101 agtgacaaaa agagtagtcc ccagaaagaa gttaagtatg aacccttttc ttttgctgat
     2161 gatattggct ccaataattg tggatactat gacttacaag cagtactaac acaccaggga
     2221 aggtctagtt cttcaggtca ttatgtatca tgggtgaaaa ggaaacaaga tgaatggatt
     2281 aagtttgatg atgacaaagt cagcatcgta acaccagaag atatcttacg gctttctggt
     2341 ggtggagact ggcatatcgc ttacgttcta ctctatgggc ctcgcagagt tgaaataatg
     2401 gaagaggaaa gtgaacagct cgagaccggt gactacaaag accatgacgg tgattataaa
     2461 gatcatgaca tcgattacaa ggatgacgat gacaagtgac cggtaagttt aaaccgctga
     2521 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2581 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2641 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2701 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2761 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2821 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2881 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2941 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3001 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3061 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3121 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggccatttcg
     3181 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3241 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3301 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3361 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3421 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3481 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3541 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggatgaccg agtacaagcc
     3601 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     3661 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     3721 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     3781 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     3841 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     3901 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     3961 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4021 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     4081 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4141 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattcgaaa tgaccgacca
     4201 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4261 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4321 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4381 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4441 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4501 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4561 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4621 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4681 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4741 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4801 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4861 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4921 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4981 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5041 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5101 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5161 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5221 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5281 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5341 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5401 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     5461 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5521 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5581 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5641 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5701 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5761 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5821 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5881 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5941 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6001 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6061 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6121 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6181 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6241 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6301 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6361 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6421 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6481 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6541 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6601 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6661 tgacgtc
//
