LOCUS       V027082                 8160 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027082
VERSION     V027082
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..2840
                     /label="FSHR(NM_000145)"
                     /note="FSHR(NM_000145)"
                     /gene="FSHR"
     CDS             2847..2912
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2936..3524
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3590..3638
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3840..4268
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4282..4611
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4678..5472
                     /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    5648..5769
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5818..5834)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5842..5858
                     /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(5866..5896)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5911..5932
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6220..6808)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6979..7839)
                     /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(7840..7944)
                     /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 ccaccatggc cctgctcctg gtctctttgc
      781 tggcattcct gagcttgggc tcaggatgtc atcatcggat ctgtcactgc tctaacaggg
      841 tttttctctg ccaagagagc aaggtgacag agattccttc tgacctcccg aggaatgcca
      901 ttgaactgag gtttgtcctc accaagcttc gagtcatcca aaaaggtgca ttttcaggat
      961 ttggggacct ggagaaaata gagatctctc agaatgatgt cttggaggtg atagaggcag
     1021 atgtgttctc caaccttccc aaattacatg aaattagaat tgaaaaggcc aacaacctgc
     1081 tctacatcaa ccctgaggcc ttccagaacc ttcccaacct tcaatatctg ttaatatcca
     1141 acacaggtat taagcacctt ccagatgttc acaagattca ttctctccaa aaagttttac
     1201 ttgacattca agataacata aacatccaca caattgaaag aaattctttc gtggggctga
     1261 gctttgaaag tgtgattcta tggctgaata agaatgggat tcaagaaata cacaactgtg
     1321 cattcaatgg aacccaacta gatgagctga atctaagcga taataataat ttagaagaat
     1381 tgcctaatga tgttttccac ggagcctctg gaccagtcat tctagatatt tcaagaacaa
     1441 ggatccattc cctgcctagc tatggcttag aaaatcttaa gaagctgagg gccaggtcga
     1501 cttacaactt aaaaaagctg cctactctgg aaaagcttgt cgccctcatg gaagccagcc
     1561 tcacctatcc cagccattgc tgtgcctttg caaactggag acggcaaatc tctgagcttc
     1621 atccaatttg caacaaatct attttaaggc aagaagttga ttatatgact caggctaggg
     1681 gtcagagatc ctctctggca gaagacaatg agtccagcta cagcagagga tttgacatga
     1741 cgtacactga gtttgactat gacttatgca atgaagtggt tgacgtgacc tgctccccta
     1801 agccagatgc attcaaccca tgtgaagata tcatggggta caacatcctc agagtcctga
     1861 tatggtttat cagcatcctg gccatcactg ggaacatcat agtgctagtg atcctaacta
     1921 ccagccaata taaactcaca gtccccaggt tccttatgtg caacctggcc tttgctgatc
     1981 tctgcattgg aatctacctg ctgctcattg catcagttga tatccatacc aagagccaat
     2041 atcacaacta tgccattgac tggcaaactg gggcaggctg tgatgctgct ggctttttca
     2101 ctgtctttgc cagtgagctg tcagtctaca ctctgacagc tatcaccttg gaaagatggc
     2161 ataccatcac gcatgccatg cagctggact gcaaggtgca gctccgccat gctgccagtg
     2221 tcatggtgat gggctggatt tttgcttttg cagctgccct ctttcccatc tttggcatca
     2281 gcagctacat gaaggtgagc atctgcctgc ccatggatat tgacagccct ttgtcacagc
     2341 tgtatgtcat gtccctcctt gtgctcaatg tcctggcctt tgtggtcatc tgtggctgct
     2401 atatccacat ctacctcaca gtgcggaacc ccaacatcgt gtcctcctct agtgacacca
     2461 ggatcgccaa gcgcatggcc atgctcatct tcactgactt cctctgcatg gcacccattt
     2521 ctttctttgc catttctgcc tccctcaagg tgcccctcat cactgtgtcc aaagcaaaga
     2581 ttctgctggt tctgtttcac cccatcaact cctgtgccaa ccccttcctc tatgccatct
     2641 ttaccaaaaa ctttcgcaga gatttcttca ttctgctgag caagtgtggc tgctatgaaa
     2701 tgcaagccca aatttatagg acagaaactt catccactgt ccacaacacc catccaagga
     2761 atggccactg ctcttcagct cccagagtca ccagtggttc cacttacata cttgtccctc
     2821 taagtcattt agcccaaaac ggatccgact acaaagacca tgacggtgat tataaagatc
     2881 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2941 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3001 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3061 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3121 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3181 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3241 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3301 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3361 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3421 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3481 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3541 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3601 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3661 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3721 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3781 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3841 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3901 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3961 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4021 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4081 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4141 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4201 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4261 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4441 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4561 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4621 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4681 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4741 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4801 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4861 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4921 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4981 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5041 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5101 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5161 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5221 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5281 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5341 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5401 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5461 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5521 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5581 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5641 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5701 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5761 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5821 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5881 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5941 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6001 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6061 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6661 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6781 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6901 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6961 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7021 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7081 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7141 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7201 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7261 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7321 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7381 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7441 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7501 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7561 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7621 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7681 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7741 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7801 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7861 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7921 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7981 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     8041 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     8101 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
