LOCUS       V022416                 6707 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022416
VERSION     V022416
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             941..2197
                     /label="BRF2(NM_018310)"
                     /note="BRF2(NM_018310)"
                     /gene="BRF2"
     CDS             2204..2269
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2307..2531
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2577..3005
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3019..3348
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3415..4209
                     /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    4383..4504
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4553..4569)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4577..4593
                     /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(4601..4631)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4646..4667
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4955..5540)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5711..6571)
                     /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(6572..6676)
                     /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 atccgccacc atgccaggca gaggccgctg
      961 cccggactgc ggctccacgg agctggtgga agactcgcac tattcgcaga gccagctggt
     1021 gtgctccgac tgcggctgcg tggtcaccga gggggtcctt accactacct tcagcgacga
     1081 gggcaatctc cgagaggtaa catattcccg aagcacaggg gaaaacgaac aagttagtcg
     1141 cagccagcaa cgaggtctcc ggcgagtgag agacctttgt cgagttctgc agttgccacc
     1201 aacatttgag gataccgcgg ttgcctacta ccaacaggca tatcggcact ctggcatccg
     1261 agcggccagg ctgcaaaaga aggaggtgtt ggttgggtgc tgcgtcttaa tcacctgccg
     1321 acagcataac tggcccctaa caatgggggc catctgcacg ctgttgtatg cagatttgga
     1381 tgtgttttct agcacttaca tgcagatagt gaagctcctg ggactggatg tgccatctct
     1441 gtgcttggca gaactggtga agacctattg cagcagcttc aaactgttcc aagcttcacc
     1501 ttctgtgcca gccaaatacg tggaagacaa agagaagatg ctgtctcgaa caatgcagtt
     1561 ggtggagctg gcaaatgaga cgtggctggt gaccgggagg catcccttgc ccgtcatcac
     1621 tgctgcgact ttcctggctt ggcagtcgct gcagcctgca gatcggcttt catgttccct
     1681 tgcccgattt tgtaaattgg caaatgtgga cctgccctac ccggcgtcct cccgcctgca
     1741 ggagctgctg gctgtgctgc tgcggatggc tgagcagctg gcctggttac gagttctgag
     1801 acttgacaaa cggtctgtgg tgaagcacat cggtgacctt ctccagcacc gccagtcact
     1861 ggtccgctct gcctttcggg atgggacagc agaagtggag acccgagaga aggagccacc
     1921 ggggtgggga caggggcaag gagaagggga ggtgggaaat aattccttag gtttacccca
     1981 ggggaagcgg ccggccagtc ctgcccttct cttgccaccc tgcatgttga agtccccgaa
     2041 gcggatctgc cctgtacccc ctgtctccac tgtcactgga gatgagaaca tttctgatag
     2101 tgaaatagaa cagtatttgc gtacccctca ggaagttagg gactttcaga gagcccaggc
     2161 tgctagacag gctgccacga gtgtccctaa ccctcccctc gaggactaca aagaccatga
     2221 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     2281 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2341 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2401 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2461 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2521 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2581 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2641 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2701 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2761 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2821 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2881 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2941 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3001 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3181 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3241 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3301 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3361 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3421 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3481 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3541 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3601 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3661 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3721 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3781 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3841 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3901 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3961 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4021 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4081 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4141 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4201 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4261 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4321 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4381 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4441 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4501 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4741 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4801 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4861 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4921 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4981 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5041 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5101 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5161 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5221 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5281 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5341 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5401 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5461 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     5521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5821 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6001 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
