LOCUS       V027351                 5981 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027351
VERSION     V027351
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"
     sig_peptide     905..964
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin&nbsp;κ
                     light chain"
                     /transl_table=1
                     /translation="METDTLLLWVLLLWVPGSTG"
                     /label="Igκ leader"
     CDS             1049..1888
                     /label="HES1(NM_005524)"
                     /note="HES1(NM_005524)"
                     /gene="HES1"
     CDS             1904..1933
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             1949..1966
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    1995..2219
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2265..2693
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2707..3036
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        3084..3131
                     /note="synthetic bacterial promoter&nbsp;"
                     /label="EM7 promoter"
     CDS             3150..3524
                     /codon_start=1
                     /gene="Sh&nbsp;ble from&nbsp;Streptoalloteichus
                     hindustanus"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin™"
                     /product="antibiotic-binding protein"
                     /transl_table=1
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD*"
                     /label="BleoR"
     polyA_signal    3654..3775
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3824..3840)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3848..3864
                     /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(3872..3902)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3917..3938
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4226..4814)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4985..5845)
                     /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(5846..5950)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 caccatggag acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac
      961 tggtgacggt tggactttga attctgctgg ttatttgttg ggtaaaatta atttgaaagc
     1021 tttggctgct ttggctaaaa aaattttgat gccagctgat ataatggaga aaaattcctc
     1081 gtccccggtg gctgctaccc cagccagtgt caacacgaca ccggataaac caaagacagc
     1141 atctgagcac agaaagtcat caaagcctat tatggagaaa agacgaagag caagaataaa
     1201 tgaaagtctg agccagctga aaacactgat tttggatgct ctgaagaaag atagctcgcg
     1261 gcattccaag ctggagaagg cggacattct ggaaatgaca gtgaagcacc tccggaacct
     1321 gcagcgggcg cagatgacgg ctgcgctgag cacagaccca agtgtgctgg ggaagtaccg
     1381 agccggcttc agcgagtgca tgaacgaggt gacccgcttc ctgtccacgt gcgagggcgt
     1441 taataccgag gtgcgcactc ggctgctcgg ccacctggcc aactgcatga cccagatcaa
     1501 tgccatgacc taccccgggc agccgcaccc cgccttgcag gcgccgccac cgcccccacc
     1561 gggacccggc ggcccccagc acgcgccgtt cgcgccgccg ccgccactcg tgcccatccc
     1621 cgggggcgcg gcgccccctc ccggcggcgc cccctgcaag ctgggcagcc aggctggaga
     1681 ggcggctaag gtgtttggag gcttccaggt ggtaccggct cccgatggcc agtttgcttt
     1741 cctcattccc aacggggcct tcgcgcacag cggccctgtc atccccgtct acaccagcaa
     1801 cagcggcacc tccgtgggcc ccaacgcagt gtcaccttcc agcggcccct cgcttacggc
     1861 ggactccatg tggaggccgt ggcggaacac tcgaggaggg cccgaacaaa aactcatctc
     1921 agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag tttaaacccg
     1981 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2041 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2101 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2161 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2221 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     2281 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2341 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2401 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     2461 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2521 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2581 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     2641 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     2701 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2761 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2821 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2881 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2941 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3001 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3061 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     3121 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     3181 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     3241 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     3301 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     3361 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     3421 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     3481 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     3541 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3601 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3661 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3721 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3781 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3841 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3901 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3961 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4021 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4081 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     4441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4681 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4741 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5821 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5941 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
