LOCUS       V043988                 8137 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043988
VERSION     V043988
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             964..3627
                     /label="HMGCR(NM_000859)"
                     /note="HMGCR(NM_000859)"
                     /gene="HMGCR"
     CDS             3634..3699
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3737..3961
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4007..4435
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4449..4778
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4845..5639
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5813..5934
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5983..5999)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6007..6023
                     /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(6031..6061)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6076..6097
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6385..6970)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7141..8001)
                     /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(8002..8106)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgttgt caagactttt tcgaatgcat ggcctctttg tggcctccca tccctgggaa
     1021 gtcatagtgg ggacagtgac actgaccatc tgcatgatgt ccatgaacat gtttactggt
     1081 aacaataaga tctgtggttg gaattatgaa tgtccaaagt ttgaagagga tgttttgagc
     1141 agtgacatta taattctgac aataacacga tgcatagcca tcctgtatat ttacttccag
     1201 ttccagaatt tacgtcaact tggatcaaaa tatattttgg gtattgctgg ccttttcaca
     1261 attttctcaa gttttgtatt cagtacagtt gtcattcact tcttagacaa agaattgaca
     1321 ggcttgaatg aagctttgcc ctttttccta cttttgattg acctttccag agcaagcaca
     1381 ttagcaaagt ttgccctcag ttccaactca caggatgaag taagggaaaa tattgctcgt
     1441 ggaatggcaa ttttaggtcc tacgtttacc ctcgatgctc ttgttgaatg tcttgtgatt
     1501 ggagttggta ccatgtcagg ggtacgtcag cttgaaatta tgtgctgctt tggctgcatg
     1561 tcagttcttg ccaactactt cgtgttcatg actttcttcc cagcttgtgt gtccttggta
     1621 ttagagcttt ctcgggaaag ccgcgagggt cgtccaattt ggcagctcag ccattttgcc
     1681 cgagttttag aagaagaaga aaataagccg aatcctgtaa ctcagagggt caagatgatt
     1741 atgtctctag gcttggttct tgttcatgct cacagtcgct ggatagctga tccttctcct
     1801 caaaacagta cagcagatac ttctaaggtt tcattaggac tggatgaaaa tgtgtccaag
     1861 agaattgaac caagtgtttc cctctggcag ttttatctct ctaaaatgat cagcatggat
     1921 attgaacaag ttattaccct aagtttagct ctccttctgg ctgtcaagta catcttcttt
     1981 gaacaaacag agacagaatc tacactctca ttaaaaaacc ctatcacatc tcctgtagtg
     2041 acacaaaaga aagtcccaga caattgttgt agacgtgaac ctatgctggt cagaaataac
     2101 cagaaatgtg attcagtaga ggaagagaca gggataaacc gagaaagaaa agttgaggtt
     2161 ataaaaccct tagtggctga aacagatacc ccaaacagag ctacatttgt ggttggtaac
     2221 tcctccttac tcgatacttc atcagtactg gtgacacagg aacctgaaat tgaacttccc
     2281 agggaacctc ggcctaatga agaatgtcta cagatacttg ggaatgcaga gaaaggtgca
     2341 aaattcctta gtgatgctga gatcatccag ttagtcaatg ctaagcatat cccagcctac
     2401 aagttggaaa ctctgatgga aactcatgag cgtggtgtat ctattcgccg acagttactt
     2461 tccaagaagc tttcagaacc ttcttctctc cagtacctac cttacaggga ttataattac
     2521 tccttggtga tgggagcttg ttgtgagaat gttattggat atatgcccat ccctgttgga
     2581 gtggcaggac ccctttgctt agatgaaaaa gaatttcagg ttccaatggc aacaacagaa
     2641 ggttgtcttg tggccagcac caatagaggc tgcagagcaa taggtcttgg tggaggtgcc
     2701 agcagccgag tccttgcaga tgggatgact cgtggcccag ttgtgcgtct tccacgtgct
     2761 tgtgactctg cagaagtgaa agcctggctc gaaacatctg aagggttcgc agtgataaag
     2821 gaggcatttg acagcactag cagatttgca cgtctacaga aacttcatac aagtatagct
     2881 ggacgcaacc tttatatccg tttccagtcc aggtcagggg atgccatggg gatgaacatg
     2941 atttcaaagg gtacagagaa agcactttca aaacttcacg agtatttccc tgaaatgcag
     3001 attctagccg ttagtggtaa ctattgtact gacaagaaac ctgctgctat aaattggata
     3061 gagggaagag gaaaatctgt tgtttgtgaa gctgtcattc cagccaaggt tgtcagagaa
     3121 gtattaaaga ctaccacaga ggctatgatt gaggtcaaca ttaacaagaa tttagtgggc
     3181 tctgccatgg ctgggagcat aggaggctac aacgcccatg cagcaaacat tgtcaccgcc
     3241 atctacattg cctgtggaca ggatgcagca cagaatgttg gtagttcaaa ctgtattact
     3301 ttaatggaag caagtggtcc cacaaatgaa gatttatata tcagctgcac catgccatct
     3361 atagagatag gaacggtggg tggtgggacc aacctactac ctcagcaagc ctgtttgcag
     3421 atgctaggtg ttcaaggagc atgcaaagat aatcctgggg aaaatgcccg gcagcttgcc
     3481 cgaattgtgt gtgggaccgt aatggctggg gaattgtcac ttatggcagc attggcagca
     3541 ggacatcttg tcaaaagtca catgattcac aacaggtcga agatcaattt acaagacctc
     3601 caaggagctt gcaccaagaa gacagccctc gaggactaca aagaccatga cggtgattat
     3661 aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc cgtttaaacc
     3721 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3781 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3841 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3901 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3961 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4021 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4081 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4141 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4201 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4261 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4321 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4381 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4441 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4501 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4561 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4621 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4681 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4741 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4801 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4861 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4921 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4981 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5041 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5101 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5161 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5221 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5281 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5341 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5401 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5461 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5521 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5581 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5641 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5701 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5761 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5821 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5881 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5941 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6001 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6061 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6121 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6181 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6901 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8101 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
