LOCUS       V020481                 8846 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020481
VERSION     V020481
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             941..4336
                     /label="DAB2IP(NM_032552)"
                     /note="DAB2IP(NM_032552)"
                     /gene="DAB2IP"
     CDS             4343..4408
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4446..4670
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4716..5144
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5158..5487
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5554..6348
                     /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    6522..6643
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6692..6708)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6716..6732
                     /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(6740..6770)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6785..6806
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7094..7679)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7850..8710)
                     /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(8711..8815)
                     /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 atccgccacc atggagcccg actcccttct
      961 ggaccaagac gactcctacg agtcgcctca agaaaggccg ggctctcggc gcagcctgcc
     1021 tggcagcctt tccgagaaga gccccagcat ggagccctcg gccgccacgc cgttccgggt
     1081 cacgggcttc ctcagccgcc gcctcaaggg ctccatcaag cgcaccaaga gccagcccaa
     1141 gctggaccgc aaccacagct tccgccacat cctgccgggg ttccggagcg ccgccgccgc
     1201 cgccgcggac aatgagaggt cccatctgat gccgaggctg aaggagtctc gctcccacga
     1261 gtccctgctc agccccagca gtgcggtgga ggcgctggac ctcagcatgg aggaagaggt
     1321 ggtcatcaag cccgtgcaca gcagcatcct tggccaggac tactgcttcg aggtgacgac
     1381 gtcatcagga agcaagtgct tttcctgccg gtctgcagct gagcgggata agtggatgga
     1441 gaacctccgg cgagcggtgc atcccaacaa ggacaacagc cggcgtgtgg agcacatcct
     1501 gaagctgtgg gtgatcgagg ccaaggacct gccagccaag aagaagtacc tgtgcgagct
     1561 gtgcctggac gatgtgctct atgcccgcac cacgggcaag ctcaagacgg acaatgtttt
     1621 ctggggcgag cacttcgagt tccacaactt gccgcctctg cgcacggtca ctgtccacct
     1681 gtaccgggag accgacaaga agaagaagaa ggagcgcaac agttacctgg gcctggtgag
     1741 cctacctgct gcctcggtgg ccgggcggca gttcgtggag aagtggtacc cggtggtgac
     1801 gcccaacccc aagggcggca agggccctgg acccatgatc cgcatcaagg cgcgctacca
     1861 aaccatcacc atcctgccca tggagatgta caaagagttc gctgagcaca tcaccaacca
     1921 ctacctgggg ctgtgtgcag ccctcgagcc catcctcagt gccaagacca aggaggagat
     1981 ggcatctgcc ctggtgcaca tcctgcagag cacgggcaag gtgaaggact tcctgacaga
     2041 cctgatgatg tcagaggtgg accgctgcgg ggacaacgag cacctcatct tccgggagaa
     2101 cacactggcc accaaggcca ttgaggagta cctcaagcta gtgggccaga agtacctgca
     2161 ggacgcccta ggtgagttca tcaaagcgct gtatgagtca gatgagaact gcgaagtgga
     2221 tcccagcaag tgctcggccg ctgacctccc agagcaccag ggcaacctca agatgtgctg
     2281 cgagctggcc ttctgcaaga tcatcaactc ctactgtgtc ttcccacggg agttgaaaga
     2341 ggtgtttgcc tcgtggaggc aggagtgcag cagtcgcggc cgcccggaca tcagtgagcg
     2401 gctcatcagc gcctccctct tcctgcgctt cctctgccca gccatcatgt cgccctcact
     2461 cttcaacctg ctgcaggagt accctgatga ccgcactgcc cgcaccctca ccctcatcgc
     2521 caaggtcacc cagaacctgg ccaactttgc caaatttggc agcaaggagg aatacatgtc
     2581 cttcatgaac cagttcctag agcatgagtg gaccaacatg cagcgcttcc tgctggagat
     2641 ctccaacccc gagaccctct ccaatacagc cggcttcgag ggctacatcg acctgggccg
     2701 cgagctctcc agcctgcact cactgctctg ggaggccgtc agccagctgg agcagagcat
     2761 agtatccaaa ctgggacccc tgcctcggat cctgagggac gtccacacag cactgagcac
     2821 cccaggtagc gggcagctcc cagggaccaa tgacctggcc tccacaccgg gctctggcag
     2881 cagcagcatc tcagctgggc tgcagaagat ggtgattgag aacgatcttt ccggtctgat
     2941 agatttcacc cggttaccgt ctccaacccc cgaaaacaag gacttgtttt ttgtcacaag
     3001 gtcctccggg gtccagccct cacctgcccg cagctcgagt tactcggaag ccaacgagcc
     3061 tgatcttcag atggccaacg gtggcaagag cctctccatg gtggacctcc aggacgcccg
     3121 cacgctggat ggggaggcag gctccccggc gggccccgac gtcctcccca cagatgggca
     3181 ggccgctgca gctcagctgg tggccgggtg gccggcccgg gcaaccccag tgaacctggc
     3241 agggctggcc acggtgcggc gggcaggcca gacaccaacc acaccaggca cctccgaggg
     3301 cgcgccaggc cggccccagc tgttggcacc gctctccttc cagaaccctg tgtaccagat
     3361 ggcggctggc ctgccgctgt caccccgtgg ccttggcgac tcaggctctg agggccacag
     3421 ctccctgagc tcacacagca acagcgagga gttggcggct gctgccaagc tgggaagttt
     3481 cagcactgcc gcggaggagc tggctcggcg gcccggtgag ctggcacggc gacagatgtc
     3541 actgactgaa aaaggcgggc agcccacggt gccacggcag aacagtgctg gcccccagag
     3601 gaggatcgac cagcctccgc ccccaccccc gccgccacct cctgcccccc gcggccggac
     3661 gccccccaac ctgctgagca ccctgcagta cccaagaccc tcaagcggaa ccctggcgtc
     3721 ggcctcacct gattgggtgg gccccagtac ccgcctgagg cagcagtcct cttcctccaa
     3781 gggggacagc ccagaactga agccacgggc agtgcacaag cagggccctt cacctgtgag
     3841 ccccaatgcc ctggaccgca cagccgcttg gctcttgacc atgaacgcgc agttgttaga
     3901 agacgagggc ctgggcccag acccccccca cagggatagg ctaaggagta aggacgagct
     3961 cagccaagca gaaaaggacc tggcggtgct gcaggacaag ctgcgaatct ccaccaagaa
     4021 gctggaggag tatgagaccc tgttcaagtg ccaggaggag acgacgcaga agctggtgct
     4081 ggagtaccag gcacggctgg aggagggcga ggagcggctg cggcggcagc aggaggacaa
     4141 ggacatccag atgaagggca tcatcagcag gttgatgtcc gtggaggaag aactgaagaa
     4201 ggaccacgca gagatgcaag cggctgtgga ctccaaacag aagatcattg atgcccagga
     4261 gaagcgcatt gcctcgttgg atgccgccaa tgcccgcctc atgagtgccc tgacccagct
     4321 gaaagagagt atgcatctcg aggactacaa agaccatgac ggtgattata aagatcatga
     4381 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     4441 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4501 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4561 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4621 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4681 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4741 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4801 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4861 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4921 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4981 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5041 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5101 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     5161 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5221 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5281 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5341 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5401 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5461 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5521 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5581 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5641 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5701 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5761 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5821 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5881 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5941 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6001 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6061 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     6121 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6181 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6241 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6301 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6361 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6421 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6481 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6541 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6601 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6661 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6721 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6781 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6841 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6901 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6961 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7021 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7501 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7561 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7621 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7681 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7741 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7801 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7861 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7921 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7981 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8041 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8101 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8161 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8221 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8281 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8341 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8401 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8461 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8521 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8581 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8641 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8701 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8761 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8821 ttccccgaaa agtgccacct gacgtc
//
