LOCUS       V022678                 7420 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022678
VERSION     V022678
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             964..2910
                     /label="UBASH3B(NM_032873)"
                     /note="UBASH3B(NM_032873)"
                     /gene="UBASH3B"
     CDS             2917..2982
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3020..3244
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3290..3718
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3732..4061
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4128..4922
                     /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    5096..5217
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5266..5282)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5290..5306
                     /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(5314..5344)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5359..5380
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5668..6253)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6424..7284)
                     /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(7285..7389)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggctc agtacggcca ccccagtccg ctcggcatgg ctgcgagaga ggagctgtac
     1021 agcaaagtca ccccccggag gaaccgccaa cagcgccccg gcaccatcaa gcatggatcg
     1081 gcgctggacg tgctcctctc catggggttc cccagagccc gcgcacaaaa agccttggca
     1141 tccacgggag gaagaagtgt tcaggcagca tgtgactggt tattctccca tgtcggtgac
     1201 cccttcctgg atgaccccct gccccgggag tacgtcctct acctccgtcc caccggcccc
     1261 ttagcacaga agctttccga cttttggcag cagtcgaagc agatctgcgg gaagaacaag
     1321 gcacacaaca tcttccccca catcacactc tgccagttct ttatgtgcga ggacagcaag
     1381 gtggatgccc tgggggaagc cctgcagacc acggtcagtc gctggaaatg taagttctcg
     1441 gccccgctgc ccctggagct ctatacgtcg tccaacttca tcggcctctt tgtaaaggaa
     1501 gacagtgcgg aggtcctcaa gaagtttgct gctgactttg ctgcagaggc tgcatccaaa
     1561 accgaagtgc atgtggaacc tcataagaag cagctacatg tgaccctggc ttaccacttc
     1621 caagccagcc acctacccac cctagagaaa ctggcccaga acattgacgt caagctaggg
     1681 tgtgactggg tggctaccat attttctcgg gatatccgat ttgctaacca tgagacatta
     1741 caggtcatct acccctatac cccacaaaat gacgatgagc tggagctggt ccccggggac
     1801 ttcatcttca tgtctccaat ggagcagacc agcaccagcg agggttggat ctatggcacg
     1861 tccttaacca ccggctgctc tggactcctg cctgagaatt acattaccaa ggctgatgaa
     1921 tgcagcacct ggatatttca tggttcttat tcaatcttaa atacatcgtc atccaactct
     1981 ctcacgtttg gggatggagt attggagagg cggccttatg aggaccaggg gctcggggag
     2041 acgactcctc ttactatcat ctgccagccc atgcagccgc tgagggtcaa cagccagccc
     2101 ggcccccaga agcgatgcct ttttgtgtgt cggcatggtg agaggatgga tgttgtgttt
     2161 gggaagtact ggctgtccca gtgcttcgat gccaaaggcc gctacatacg caccaacctg
     2221 aacatgcctc atagtttacc tcagcggagt ggtggtttcc gagattacga gaaagatgct
     2281 cccatcactg tgtttggatg catgcaagca agactagtgg gtgaagcctt attagagagc
     2341 aataccatta tcgatcatgt ctattgctcc ccgtcccttc gctgcgttca gactgcacac
     2401 aatatcttga aaggtttaca acaagaaaat cacttgaaga tccgtgtaga gcccggctta
     2461 tttgagtgga caaaatgggt tgctgggagc acattacctg catggatacc tccatcagag
     2521 ttagctgcag ccaacctgag tgttgataca acctacagac ctcacattcc aatcagcaaa
     2581 ttagttgttt cagaatccta tgatacttat atcagtagaa gtttccaagt aacaaaagaa
     2641 ataataagtg aatgtaaaag taaaggaaat aacatcctga ttgtggccca cgcatcttcc
     2701 cttgaagcgt gtacctgcca acttcagggc ctgtcacctc agaactccaa ggacttcgta
     2761 caaatggtcc gaaagatccc atatctggga ttttgttcct gtgaagaatt aggagaaact
     2821 ggaatatggc agctgacaga tccaccaatc cttcctctta cccatggacc aactgggggc
     2881 ttcaactgga gagagacctt gcttcaagaa ctcgaggact acaaagacca tgacggtgat
     2941 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgtttaa
     3001 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3061 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3121 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3181 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3241 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3361 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3481 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3601 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3721 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3781 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3961 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4081 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4141 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4201 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4261 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4321 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4381 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4441 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4501 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4561 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4621 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4681 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4741 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4801 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4861 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4921 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4981 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5041 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5101 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5161 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5221 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5281 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5341 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5401 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5461 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5521 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5581 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5641 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5701 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5761 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5821 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5881 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5941 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6001 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6061 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6121 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6181 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6301 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6361 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6421 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6481 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6541 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6601 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6661 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6721 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6781 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6841 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6901 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6961 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7021 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7081 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7141 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7201 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7261 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7321 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7381 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
