LOCUS       V043804                 7627 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043804
VERSION     V043804
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..3117
                     /label="FBXL18(NM_024963)"
                     /note="FBXL18(NM_024963)"
                     /gene="FBXL18"
     CDS             3124..3189
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3227..3451
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3497..3925
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3939..4268
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4335..5129
                     /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    5303..5424
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5473..5489)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5497..5513
                     /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(5521..5551)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5566..5587
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5875..6460)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6631..7491)
                     /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(7492..7596)
                     /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 accatggcca gctccggaga ggacatatcc aatgatgatg atgacatgca ccctgcagca
     1021 gccgggatgg cagacggggt ccacctccta gggttctctg atgagatcct ccttcacatc
     1081 ctgagtcacg tccccagcac agatctgatt ctgaacgtcc ggcgtacctg tcggaagctt
     1141 gcagccctgt gccttgacaa gagcctcatc cacaccgtgt tgctgcaaaa ggactatcag
     1201 gcgagcgagg acaaagtgag gcagctggtg aaggagatcg gccgggagat ccagcagctg
     1261 agcatggctg gctgctactg gctgcctggc tccaccgtgg aacacgtggc ccgctgccgc
     1321 agcctggtga aggtgaacct ctcgggctgc cacctcactt ccctgcgcct ctccaagatg
     1381 ctctcggccc tgcagcacct gcgctcgctg gccatcgacg tgagccccgg cttcgacgcc
     1441 agccagctga gcagcgagtg caaggccacc ctgagccgcg tgcgggagct caagcagacg
     1501 ctgttcactc cctcctacgg cgtggtgccc tgctgcacca gcctagagaa gctgctgctc
     1561 tacttcgaga ttctggaccg cacgcgcgag ggcgccatcc tctcgggcca gcttatggtg
     1621 ggccagagca acgtgccgca ctaccagaac ctgcgggtct tctatgcgcg cctggccccc
     1681 ggctacatca accaggaggt ggtgcggctc tacctggctg tgcttagcga ccgcactcct
     1741 cagaacctcc acgccttcct catctccgtc cctggcagct tcgcggagag cggcgccacc
     1801 aagaacctcc tggactccat ggcgcgcaat gtcgtgctgg atgccctgca gctgcccaag
     1861 tcctggctga acggctcttc cctcctgcag cacatgaaat tcaacaaccc gttctacttc
     1921 agtttcagcc gctgtaccct gtcaggcggc catctgatcc agcaggtcat caacggcggg
     1981 aaggacctgc ggagcctggc cagcttgaac ctcagcggct gcgtccactg cctgtcccca
     2041 gactcgctgc tccgcaaggc ggaggacgac atcgacagca gcatcctgga gactctggtg
     2101 gcgtcctgct gcaacctgcg ccacctgaac ctctcggccg cccaccacca cagctcggag
     2161 ggcctgggcc gccacctctg ccagctcctg gcccggctgc gtcacctgcg ctccctctcc
     2221 ctgcctgtct gctctgtcgc tgactccgcg ccgcgcgccg accgcgcgcc cgcccagccg
     2281 gccatgcacg cagtgccgcg cggctttggc aagaaagtgc gtgtgggcgt gcagtcctgt
     2341 cccagcccct tctcgggcca ggcgtgcccc cagccctcct ccgtgttctg gtctctgctg
     2401 aagaacctgc ccttcctgga acacctcgag ctgattgggt ccaacttctc ctccgccatg
     2461 ccccgcaacg agcccgccat ccgcaactcg ctcccaccct gcagccgcgc acagagtgtc
     2521 ggggactcgg aggtggccgc catcggccag ctggccttcc tgcggcacct gacgctcgca
     2581 cagctgccca gcgtccttac gggctccggg ctggtcaata tcggcctgca gtgccagcag
     2641 ttgcggtccc tgtcgctggc caacctgggc atgatgggga aggtggtgta catgcccgcg
     2701 ctctcagaca tgttgaagca ctgcaagcgg ctgagggacc tcaggctgga gcagccctac
     2761 ttcagcgcca acgcccagtt cttccaggcg ctgagccagt gcccctcgct gcagcgcctg
     2821 tgcctggtct ctcgcagcgg caccctccag cccgatgccg tgctggcctt catggctcgc
     2881 tgcctgcagg ttgtcatgtg ccacctgttc accggggagt ccctcgccac ctgcaagagc
     2941 ctgcagcagt cgcttctccg cagcttccag gccgagcggc ccgcgttaaa cgtcgtcatc
     3001 ttccctctgc tccacgaggg cctgaccgac gtcatccggg acgtccccct ggtgcacctg
     3061 gatgagatca ccttatttaa gagcagagtg gccgaggaac cgccgaacct gtggtggctc
     3121 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3181 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3241 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3301 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3361 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3421 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3481 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3541 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3721 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3781 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3841 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3901 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4021 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4141 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4201 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4261 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4321 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4381 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4441 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4501 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4561 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4621 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4681 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4741 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4801 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4861 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4921 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4981 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5041 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5101 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5161 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5221 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5281 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5341 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5401 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5521 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7561 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7621 tgacgtc
//
