LOCUS       V039421                 7953 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039421
VERSION     V039421
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             966..3425
                     /label="DDX23(NM_004818)"
                     /note="DDX23(NM_004818)"
                     /gene="DDX23"
     CDS             3438..3467
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3483..3500
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3550..3774
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3820..4248
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4262..4591
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4658..5452
                     /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    5626..5747
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5796..5812)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5820..5836
                     /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(5844..5874)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5889..5910
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6198..6786)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6957..7817)
                     /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(7818..7922)
                     /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 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcg
      961 ccaccatggc aggagagctg gctgacaaaa aggaccgtga tgcatcacct tccaaggagg
     1021 aaaggaagcg atcacggact cctgacagag agcgggatag agaccgggac cggaagtctt
     1081 ccccatctaa agatagaaag cggcatcgtt caagggatag acgtcgagga ggcagccgtt
     1141 ctcgctctcg ttcccgttcc aaatctgcag aaagagaacg acggcacaaa gaacgagaac
     1201 gagataagga gcgggatcgg aataagaagg accgagatcg agacaaggat gggcacagac
     1261 gggacaagga ccgtaaacga tccagcttat ctcctggtcg aggaaaagac tttaaatctc
     1321 ggaaggacag agactctaag aaggatgaag aggatgaaca tggtgataag aagcctaagg
     1381 cccagccatt atccctggag gagcttctgg ccaagaaaaa ggctgaggaa gaagctgagg
     1441 ctaagcccaa gttcctctct aaagcagaac gagaggctga agctctaaag cgacggcagc
     1501 aggaggtgga agagcggcag aggatgcttg aagaagagag gaagaaaagg aaacagttcc
     1561 aagacttggg caggaagatg ttggaagatc ctcaggaacg ggaacgtcgg gaacgcaggg
     1621 agaggatgga acgggagacc aatggaaatg aggatgagga agggcggcag aagatccggg
     1681 aagagaagga taagagcaag gaactgcatg ccattaagga gcgttacctg ggtggcatca
     1741 aaaagcggcg ccgaacgaga catctcaatg accggaaatt tgtttttgag tgggatgcat
     1801 ctgaggacac atccattgac tacaaccccc tgtacaaaga acggcaccag gtgcagttgt
     1861 tagggcgagg cttcattgca ggcattgacc tcaagcagca gaagcgagag cagtcacgtt
     1921 tctatggaga cctaatggag aagaggcgaa ccctggaaga aaaggagcag gaggaggcaa
     1981 gactccgcaa acttcgtaag aaggaagcca agcagcgctg ggatgatcgt cattggtctc
     2041 agaaaaagtt agatgagatg acggacaggg actggcggat cttccgtgag gactacagca
     2101 tcaccaccaa aggtggcaag atccccaatc ccatccgatc ctggaaagac tcttctctgc
     2161 ccccacacat cttggaggtc attgataagt gtggctacaa ggaaccaaca cctatacagc
     2221 gtcaggcaat tcccattggg ctacagaatc gtgacatcat tggtgtggct gagactggca
     2281 gtggcaagac agcagccttc ctcatccctc tgctggtctg gatcaccaca cttcccaaaa
     2341 ttgacaggat cgaagagtca gaccaaggcc cttatgccat catcctggct cccacccgtg
     2401 agttggctca acagattgag gaagagacca tcaagtttgg gaaaccgcta ggtatccgca
     2461 ctgtggctgt cattggtggc atctccagag aagaccaggg cttcaggctg cgcatgggtt
     2521 gtgagattgt gattgctacc cctgggcgtt tgattgatgt gctggagaac cgctacctgg
     2581 tgctgagccg ctgtacctat gtggttctgg atgaggcaga taggatgatt gacatgggct
     2641 ttgagccaga tgtccagaag atcctggagc acatgcctgt cagcaaccag aagccagaca
     2701 cggatgaggc tgaggaccct gagaagatgc tggccaactt tgagtcggga aaacataagt
     2761 accgccaaac agtcatgttc acggccacca tgcccccagc ggtggagcgt ctggccagga
     2821 gctatcttcg gcgacctgct gtggtgtaca ttggctccgc aggcaagccc catgagcgtg
     2881 tggaacagaa ggtcttcctc atgtcagagt cagaaaagag gaaaaagctg ctggcaatct
     2941 tggagcaagg ctttgaccca cccatcatta tttttgtcaa ccagaagaag ggctgcgacg
     3001 tgttggccaa atccctggag aagatggggt acaatgcttg cacactgcac ggtggaaaag
     3061 gccaggagca gcgagagttt gcgttgtcca acctcaaggc tggggccaag gatattttgg
     3121 tggctacaga tgtggctggt cgtggtattg acatccaaga tgtgtctatg gttgtcaact
     3181 atgatatggc caaaaatatt gaagattaca tccaccgcat tggccgcacg ggacgagcag
     3241 gcaagagtgg ggtggccatc accttcctca caaaagagga ctctgctgtg ttctacgagc
     3301 tgaagcaagc tatcctggaa agcccagtgt cttcctgtcc ccccgaacta gccaaccacc
     3361 cagatgccca gcataagcca ggcaccatcc tcaccaagaa gcgccgggaa gagaccatct
     3421 ttgccaagct tgggcccgaa caaaaactca tctcagaaga ggatctgaat agcgccgtcg
     3481 accatcatca tcatcatcat tgagtttaaa cggtctccag cttaagttta aaccgctgat
     3541 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3601 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3661 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3721 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3781 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4021 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4141 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4201 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4261 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4321 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4441 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4501 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4561 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4621 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4681 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4741 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4801 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4861 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4921 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4981 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5041 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5101 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5161 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5221 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5281 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5341 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5401 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5461 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5521 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5581 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5641 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5701 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5761 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5821 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5881 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5941 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6001 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6061 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6121 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6601 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6961 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7021 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7081 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7141 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7201 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7261 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7321 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7381 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7441 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7501 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7561 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7621 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7681 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7741 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7801 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7861 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7921 cgcacatttc cccgaaaagt gccacctgac gtc
//
