LOCUS       V023830                 7988 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023830
VERSION     V023830
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             782..808
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             812..838
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             842..868
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             878..2668
                     /label="FLCN(NM_001353229)"
                     /note="FLCN(NM_001353229)"
                     /gene="FLCN"
     CDS             2675..2740
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2764..3352
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3418..3466
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3668..4096
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4110..4439
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4506..5300
                     /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    5476..5597
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5646..5662)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5670..5686
                     /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(5694..5724)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5739..5760
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6048..6636)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6807..7667)
                     /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(7668..7772)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 gtacccatac gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag
      841 ctatccttat gatgtcccag actacgcatg cggccgcatg aatgccatcg tggctctctg
      901 ccacttctgc gagctccacg gcccccgcac tctcttctgc acggaggtgc tgcacgcccc
      961 acttcctcaa ggggatggga atgaggacag tcctggccag ggtgagcagg cggaagaaga
     1021 ggaaggtggc attcagatga acagtcggat gcgtgcgcac agccccgcag agggggccag
     1081 cgtcgagtcc agcagcccgg ggcccaaaaa gtcggacatg tgcgagggct gccggtcact
     1141 tgctgcaggg cacccgggat atatcagcca tgataaagag acctccatta aatacgtcag
     1201 ccaccagcac cccagccacc cccagctctt cagcattgtc cgccaggcct gtgtccggag
     1261 cctgagctgt gagtgcaggg agagtgcagt ggctgctctg acctgctggt tcttctgcat
     1321 gctccaggtc tgccctggcc gtgaaggccc catcttcttc ggagatgagc agcacggctt
     1381 tgtgttcagc cacaccttct tcatcaagga cagcctggcc aggggcttcc agcgctggta
     1441 cagcatcatc accatcatga tggaccggat ctacctcatc aactcctggc ccttcctgct
     1501 ggggaaggtc cggggaatca tcgatgagct ccagggcaag gcgctcaagg tgtttgaggc
     1561 agagcagttt ggatgcccac agcgtgctca gaggatgaac acagccttca cgccattcct
     1621 acaccagagg aacggcaacg ccgcccgctc gctgacatcg ctgacaagtg atgacaacct
     1681 gtgggcgtgc ctgcacacct cctttgcctg gctcctgaag gcgtgtggca gccggctgac
     1741 cgagaagctc ctggaaggtg ctccgaccga ggataccttg gtccagatgg agaagctcgc
     1801 tgatttagaa gaggaatcag aaagctggga caactctgag gctgaagagg aggagaaagc
     1861 ccctgtgttg ccagagagta cagaagggcg ggagctgacc cagggcccgg cagagtcctc
     1921 ctctctctca ggctgtggga gctggcagcc ccggaagctg ccagtcttca agtccctccg
     1981 gcacatgagg caggtcctgg gtgccccttc tttccgcatg ctggcctggc acgttctcat
     2041 ggggaaccag gtgatctgga aaagcagaga cgtggacctc gtccagtcag cttttgaagt
     2101 acttcggacc atgcttcccg tgggctgcgt ccgcatcatc ccatacagca gccagtacga
     2161 ggaggcctat cggtgcaact tcctggggct cagcccgcac gtgcagatcc ccccccacgt
     2221 gctctcctca gagtttgctg tcatcgtgga ggtccacgca gccgcacgtt ccaccctcca
     2281 ccctgtgggg tgtgaggatg accagtctct cagcaagtac gagtttgtgg tgaccagtgg
     2341 gagccctgta gctgcagacc gagtgggccc caccatcctg aataagattg aagcggctct
     2401 gaccaaccag aacctgtctg tggatgtggt ggaccagtgc ctcgtctgcc tcaaggagga
     2461 gtggatgaac aaagtgaagg tgctttttaa gttcaccaag gtggacagtc gacccaaaga
     2521 ggacacacag aagctgctga gcatcctggg tgcgtccgag gaggacaatg tcaagctgct
     2581 gaagttctgg atgactggcc tgagcaagac ctacaagtca cacctcatgt ccacggtccg
     2641 cagccccaca gcctcggagt ctcggaacct cgaggactac aaagaccatg acggtgatta
     2701 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     2761 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2821 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2881 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2941 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3001 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3061 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3121 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     3181 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3241 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3301 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     3361 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3421 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3481 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3541 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3601 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     3661 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3721 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3781 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     3841 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3901 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3961 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4021 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4081 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4141 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4201 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4261 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4321 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4381 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4441 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4501 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4561 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4621 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4681 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4741 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4801 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4861 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4921 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4981 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5041 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5101 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5161 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5221 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5281 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5341 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5401 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5461 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5521 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5581 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5641 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5701 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5761 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5821 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5881 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5941 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6001 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6241 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6481 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7801 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     7861 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7921 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     7981 tgcttagg
//
