LOCUS       V030488                 7590 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030488
VERSION     V030488
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2270
                     /label="KLHL42(NM_020782)"
                     /note="KLHL42(NM_020782)"
                     /gene="KLHL42"
     CDS             2277..2342
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2366..2954
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3020..3068
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3270..3698
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3712..4041
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4108..4902
                     /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    5078..5199
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5248..5264)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5272..5288
                     /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(5296..5326)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5341..5362
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5650..6238)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6409..7269)
                     /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(7270..7374)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtc ggccgaggag atggtgcaga
      781 tccgcctgga ggaccgctgc tacccggtga gcaagaggaa gctcatcgag cagagcgact
      841 acttccgcgc cctctaccgc tccggcatgc gcgaggccct gagccaggag gccggcggcc
      901 cggaggtgca gcagctgcgc ggcctcagcg cgccgggcct gcggctggtg ctggacttca
      961 tcaacgccgg cggggcccgc gaaggctggc tcctgggccc gcgcggggaa aagggcggcg
     1021 gggtggacga ggacgaggag atggatgagg tgagcctgct gtccgagctg gtggaggcgg
     1081 cctccttcct gcaggtcacg tccctgctgc agctgctgct gtcccaggtg cggctcaata
     1141 actgcctgga gatgtaccgc ctggcgcagg tgtacgggct gcccgacctg caggaggcct
     1201 gcctgcgctt catggtcgtc cacttccacg aggtgctgtg caagccccag ttccacctcc
     1261 tggggtctcc tccccaagct ccaggggatg tcagcctgaa gcagaggctg agggaggccc
     1321 ggatgactgg gactcctgtc ctcgtggccc tcggggactt cctgggggga cccctggccc
     1381 ctcaccccta ccagggggag cccccgtcca tgctcaggta cgaggagatg actgagcgtt
     1441 ggttcccgct ggccaacaac cttcctcccg acctggtcaa tgtcaggggc tatgggtctg
     1501 ccatcctgga caactacctc ttcatagtgg gcgggtacag gatcactagc caggagatct
     1561 ccgctgcgca ttcctacaac cccagcacca acgagtggct ccaggtggcc tccatgaacc
     1621 agaagaggtc taacttcaaa cttgtggctg ttaattcaaa actctatgcc atcggagggc
     1681 aggccgtttc taacgttgag tgttacaacc ccgagcagga tgcgtggaat tttgtggcgc
     1741 ccttacccaa tcctctggct gagttctctg cctgtgagtg taagggaaaa atttatgtca
     1801 ttggaggata cactaccaga gaccggaaca tgaacatttt gcagtactgc ccctcttccg
     1861 acatgtggac gctctttgaa acatgtgacg tccacattcg caagcagcag atggtgtctg
     1921 tggaagagac catctacatc gtgggggggt gtctccacga gctggggccc aaccgcagga
     1981 gcagccagag cgaggacatg ctcaccgtgc agtcctacaa caccgtcacc cgccagtggc
     2041 tctacctcaa ggagaacacg tccaaatcgg gtcttaactt gacttgtgcg ctccataacg
     2101 acggcatcta catcatgagc agagacgtca ccctgtcgac cagcttggaa caccgagtgt
     2161 tcctcaagta caacatcttt tcagatagtt gggaagcatt tcggcgtttt ccagcttttg
     2221 gacataactt gctggtttct tctctttatc tgcccaataa agcagaaaca ggatccgact
     2281 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     2341 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     2401 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2461 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2521 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2581 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2641 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2701 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2761 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2821 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2881 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2941 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     3001 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     3061 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3121 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3181 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     3241 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3301 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3361 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3421 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3481 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3541 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3601 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     3661 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3721 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3781 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3841 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3901 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3961 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4021 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4081 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4141 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4201 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4261 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4321 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4381 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4441 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4501 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4561 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4621 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4681 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4741 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4801 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4861 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4921 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4981 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5041 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5101 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5161 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5221 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5281 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5341 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5401 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5461 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5521 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5581 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5641 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5701 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5761 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5821 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5881 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5941 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6001 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6061 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6121 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6181 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6241 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6301 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6361 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6421 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6481 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6541 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6601 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6661 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6721 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6781 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6841 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6901 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6961 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7021 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7081 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7141 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7201 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7261 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7321 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7381 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     7441 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7501 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7561 accgacaatt gcatgaagaa tctgcttagg
//
