LOCUS       V024061                 8380 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024061
VERSION     V024061
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             937..4131
                     /label="LRIG2(NM_014813)"
                     /note="LRIG2(NM_014813)"
                     /gene="LRIG2"
     CDS             4144..4209
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4244..4468
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4514..4942
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4956..5285
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5297..5896
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    6056..6177
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6226..6242)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6250..6266
                     /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(6274..6304)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6319..6340
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6628..7213)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7384..8244)
                     /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(8245..8349)
                     /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 aagctttcta gagaattcga tatcggatcc gccaccatgg cgccggcgcc cctaggcgtc
      961 ccggaggagc agttgctggg gtgtcgatct agagtgcttt ctcggttact cttcattgcc
     1021 cagaccgctc tcctcctgtt gcccgccgcc ggagcaggtc tctgccccgc gccctgctcc
     1081 tgccgcattc ctctcctgga ctgcagtcgc aggaaattgc ccgcaccgag ctggagggcg
     1141 ctgtcgggct tgctgccccc cgacaccgct atcctggatt tcagtcataa tcggttgtct
     1201 aactggaaca tcagcttgga atcacaaaca ttacaggaag tgaaaatgaa ttacaatgaa
     1261 ctaacagaaa tcccgtattt tggagaacct acatctaata ttactctact ttcattagtc
     1321 cataatataa tcccagaaat aaatgcacag gcactccagt tttaccctgc tctggagagt
     1381 ttagacctca gctcaaatat aatatcagaa atcaagacat cttcatttcc tcgcatgcag
     1441 cttaaatacc tgaatttaag taataacaga ataaccacct tggaggctgg ttgcttcgat
     1501 aatttatcaa gttccttatt agtggtaaag ttaaaccgta accgaatgag catgattcca
     1561 cctaagatct tcaagctgcc tcacctccaa ttcttggaac ttaaaagaaa cagaattaaa
     1621 attgtggaag gtcttacatt ccaagggctt gactccttaa gatctttgaa aatgcagcgg
     1681 aatggaatta gcaaacttaa ggatggagca ttttttggct tgaataacat ggaagaacta
     1741 gaactggaac acaacaacct tacacgagta aacaaggggt ggttgtatgg cttgcgaatg
     1801 ttacagcagc tctatgtgag ccagaatgct attgaaagaa tcagccctga tgcatgggag
     1861 ttctgccaaa gactatccga acttgatttg tcctataacc agctgacccg cctggatgaa
     1921 tctgcctttg tgggtctgag cttattggag agattgaatt taggagacaa cagagtcact
     1981 catattgctg atggtgtatt tagatttctt tccaatcttc agacattaga cttaagaaac
     2041 aatgaaattt catgggccat agaagatgct agtgaagcct ttgctggact cacaagtctc
     2101 actaaactaa tcttacaagg aaaccagatt aagtcaatta caaagaaagc attcattggt
     2161 cttgaatccc ttgagcatct agatttgaac aataatgcta taatgtctat ccaagaaaat
     2221 gctttttctc agactcattt aaaagaactg attctgaaca caagcagttt gctctgtgac
     2281 tgccatttga agtggctact tcaatggttg gttgataata actttcaaca ttctgtgaat
     2341 gtaagctgtg cacaccctga atggctagca gggcaaagca tcctgaatgt ggatctgaaa
     2401 gattttgtct gtgatgattt tctcaagcca cagataagga cacatcctga aaccataatt
     2461 gctctaagag gcatgaatgt gactctgacg tgcactgcag tgagcagcag tgattcaccc
     2521 atgtccactg tgtggcgcaa agacagtgaa atcctgtatg acgtggatac tgagaatttt
     2581 gttcgttatt ggcagcaagc tggagaagct ctggaatata ctagtatctt acatcttttc
     2641 aatgtgaatt tcacagatga aggaaaatat cagtgtattg ttactaatca ctttggttct
     2701 aattattctc agaaagccaa actgactgta aatgagatgc catcttttct gaaaacgcca
     2761 atggatctga ctattcgcac tggtgccatg gccagattag aatgtgctgc agagggacac
     2821 cctgcacctc agatttcctg gcagaaagat ggtggtactg actttcctgc ggctcgagaa
     2881 agacgcatgc acgtcatgcc cgaggatgac gtcttcttta ttgccaatgt gaaaatagaa
     2941 gatatgggaa tctatagctg catggcacaa aatacagcag gaggtctctc agcaaatgct
     3001 tccctaacag tgttagagac accctcattt attagacccc tggaggataa gacagtaaca
     3061 cgaggtgaaa ctgcggtgtt acagtgcata gctggaggga gtcctgcccc tcgtctcaac
     3121 tggactaaag atgatgggcc tttgctggtg acagaacgac atttctttgc tgcagccaat
     3181 cagcttctca tcattgtaga tgccgggcta gaagatgctg ggaaatatac ctgcattatg
     3241 tctaacaccc ttgggacaga acgtggccac atttacctaa atgtcatttc atcccccaat
     3301 tgtgactctt cccagagtag cattgggcat gaagatgatg gctggaccac agttggcatt
     3361 gtcatcattg ttgtggtctg ctgtgttgtt ggcacttctt tgatctgggt cattgttatt
     3421 taccacatga gaaggaaaaa tgaagactat agtatcacaa acacagagga gctcaatctg
     3481 cctgcagaca ttcccagcta cttgtcttcc caaggaacgc tgtctgagcc acaggaaggc
     3541 tacagcaact ctgaggcagg cagtcatcag caacttatgc ctcctgccaa tggatatata
     3601 cacaaaggca ctgacggtgg cactggtacc cgggtgattt gctcagattg ttatgacaat
     3661 gccaacatct actccaggac ccgagaatac tgtccataca cctatattgc tgaggaggac
     3721 gttcttgatc agacactgtc cagcctcatg gtccaaatgc ctaaagagac atatttagta
     3781 catcctcccc aggatactac tgccctagag agcctgatac cgtcagccaa cagagagcca
     3841 tctgcctttc ccaccaacca tgagaggata agtgagaaga aacttccctc cacacagatg
     3901 agcggtgaaa cattgcagcg gcccgtgtgg aacataaaca gagaactagg cctgcctcat
     3961 cctccttttt cccagcagcc agtccatgag tcaccacaac ttcatcaaaa tgagggcctg
     4021 gcagggagag agccagactg ttctgcttct tccatgtcct gccacaggtt acaggatcat
     4081 gcttttgatt ttagtaggac tcggaacatt caagatggta gtgagggcac actcgagacc
     4141 ggtgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
     4201 gatgacaagt gaccggtaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc
     4261 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4321 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4381 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4441 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4501 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4561 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4621 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4681 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4741 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4801 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4861 tcttttgatt tataagggat tttggccatt tcggcctatt ggttaaaaaa tgagctgatt
     4921 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4981 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5041 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5101 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5161 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5221 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5281 gcaaaaagct cccgggatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     5341 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     5401 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     5461 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5521 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5581 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5641 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5701 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5761 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5821 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5881 caagcccggt gcctgattcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5941 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6001 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6061 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6121 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6181 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6241 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6301 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6361 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6421 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6481 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6541 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7141 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
