LOCUS       V036432                 7980 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036432
VERSION     V036432
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..2660
                     /label="Rad21(NM_009009)"
                     /note="Rad21(NM_009009)"
                     /gene="Rad21"
     CDS             2667..2732
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2756..3344
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3410..3458
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3660..4088
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4102..4431
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4498..5292
                     /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    5468..5589
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5638..5654)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5662..5678
                     /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(5686..5716)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5731..5752
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6040..6628)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6799..7659)
                     /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(7660..7764)
                     /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 ccaccatgtt ctacgcacat tttgtcctca
      781 gtaaacgagg gcctctggcc aaaatctggc tggcggccca ttgggacaag aagctaacca
      841 aagcccatgt atttgagtgc aacttagaga gcagtgtgga gagcatcatc tcaccaaagg
      901 tgaagatggc gctgcggacg tcaggacacc ttctcctggg agtagtccgc atctatcaca
      961 ggaaagccaa atacctcctc gcagactgta atgaagcatt tattaaaata aagatggcgt
     1021 ttcggccagg tgttgtcgat ctacctgagg aaaatcggga agcagcttat aatgccatta
     1081 ctttacctga ggaattccac gattttgatc agccactgcc agatttagat gatattgacg
     1141 tcgcccagca gttcagcctg aaccaaagca gagtagaaga gataaccatg agagaagaag
     1201 tcggaaacat cagtatccta caggaaaatg actttggtga cttcggaatg gatgaccgtg
     1261 aaataatgag agaaggcagt gctttcgagg atgacgacat gttagtgagc accagcgctt
     1321 ccaaccttct cctcgagcca gagcagagca ccagcaacct gaatgaaaag atgaatcact
     1381 tagagtacga agaccagtac aaagatgaca attttggaga aggaaatgat ggcggtatat
     1441 tagatgacaa acttataagt aataatgatg gtggcatctt tgacgatccc cctgccttgt
     1501 ctgaggcagg ggtcatgttg ccagagcaac ctgcacatga tgacatggat gaagatgaca
     1561 atggctcact gggtgggccg gatagtcccg actctgtgga tcctgtcgaa ccgatgccaa
     1621 ctatgactga tcagacaact ctcgtcccaa acgaggaaga agcttttgcg ttggagccca
     1681 ttgatataac tgtcaaagag acaaaagcca agaggaagag gaagctgatt gttgacagtg
     1741 tcaaagaatt ggatagtaag accattagag cccagcttag cgattattct gatattgtta
     1801 cgactctgga cctggctccg ccaaccaaga agcttatgat gtggaaagag acaggaggag
     1861 tggaaaagct cttcttttta ccagcacagc ccctgtggaa taaccggcta ctgaagctct
     1921 tcacacgctg ccttacccca cttgtaccag aagaccttag gaagagaagg aaagggggag
     1981 aggcagataa tctggatgag ttcctcaaag agtttgagaa tccagaggtt cccagagagg
     2041 agcagcagcc acagcagcag cagccacagc cgcagcgaga tgtcatcgat gagcccatta
     2101 tagaagagcc aagccgcctc caggactcag tgatggaggc cagcagaaca accatagaag
     2161 aatcagccat gcccccacca ccccctcaag gagttaagcg gaaagccggg caaatagacc
     2221 cagagccttc gatacctcct cagcaggtag agcaaatgga aataccacca gtagaacttc
     2281 ccccagagga gcctccaaat atctgtcagc tgatcccgga gttagagctc ctaccggaga
     2341 aggagaagga aaaagagaag gagaaggaag aggaggagga ggaggaggat gaagatgctt
     2401 cagggggtga tcaggatcaa gaggaaagga gatggaacaa acgcactcag cagatgcttc
     2461 atggtcttca gcgagctctt gctaaaactg gagcagagtc tatcagtttg cttgagctgt
     2521 gtcgaaacac aaaccgaaag caggcagcag caaagttcta cagctttttg gttcttaaga
     2581 agcagcaagc catcgagctc acacaggaag agccgtacag tgacatcatt gcaacccctg
     2641 gaccacggtt ccatattatc ggatccgact acaaagacca tgacggtgat tataaagatc
     2701 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2761 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2821 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2881 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2941 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3001 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3061 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3121 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3181 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3241 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3301 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3361 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3421 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3481 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3541 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3601 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3661 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3721 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3781 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3841 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3901 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3961 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4021 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4081 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4261 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4321 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4381 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4441 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4501 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4561 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4621 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4681 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4741 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4801 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4861 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4921 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4981 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5041 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5101 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5161 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5221 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5281 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5341 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5401 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5461 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5581 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5641 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5701 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5761 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5821 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5881 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5941 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6001 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6061 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6121 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6181 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6241 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6301 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6361 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6421 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6481 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6541 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6601 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6661 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6721 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6781 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6841 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6901 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6961 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7021 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7081 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7141 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7201 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7261 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7321 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7381 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7441 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7501 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7561 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7621 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7801 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7861 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7921 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
