LOCUS       V026844                 7064 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026844
VERSION     V026844
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..2554
                     /label="AJUBA(NM_032876)"
                     /note="AJUBA(NM_032876)"
                     /gene="AJUBA"
     CDS             2561..2626
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2664..2888
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2934..3362
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3376..3705
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3772..4566
                     /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    4740..4861
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4910..4926)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4934..4950
                     /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(4958..4988)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5003..5024
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5312..5897)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6068..6928)
                     /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(6929..7033)
                     /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 atggagcggt taggagagaa
      961 agccagtcgc ctgctggaga agttcggccg cagaaagggt gaatctagcc ggtctgggtc
     1021 tgacgggacc cccgggccgg gcaaggggcg cctaagtggg ttggggggac ctaggaagtc
     1081 agggccccga ggagctactg ggggacctgg ggatgagccg ttggagccgg cccgggagca
     1141 aggttccctg gacgctgagc gaaatcagcg cggctccttt gaggcgccgc gctacgaagg
     1201 ctcttttccc gcggggccgc cgcccacccg ggccttgcct ctacctcagt cgttgccccc
     1261 cgattttcgg ctggagccca cggccccggc cctcagcccc cgctctagct tcgccagtag
     1321 ctcggccagc gacgcgagca agccgtccag cccccggggc agcctgctgc tggacggggc
     1381 gggggctggc ggagctggag gtagccggcc ctgcagcaat cgcaccagcg gcatcagcat
     1441 gggctacgac cagcgccacg ggagcccctt gccagcgggg ccgtgcctgt ttggcccacc
     1501 cctggccgga gcaccggcag gctattctcc cggaggggtc ccgtccgcct acccggagct
     1561 ccacgccgcc ctggaccgat tgtacgctca gcggcccgcg gggttcggct gccaggaaag
     1621 ccgccactcg tatcccccgg ccctgggcag ccctggagct ctagccgggg ccggagtggg
     1681 agcggcgggg cccttggaga gacggggggc gcaacccgga cgacactctg tgaccggcta
     1741 cggggactgc gccgtgggcg cccggtacca ggacgagcta acagctttgc ttcgcctgac
     1801 ggtgggcacc ggtgggcgag aagccggagc ccgcggagaa ccctcgggga ttgagccgtc
     1861 gggtctggag gagccaccag gtcctttcgt tccggaggcc gcccgggccc ggatgcggga
     1921 gccagaggcc agggaggact acttcggcac ctgtatcaag tgcaacaaag gcatctatgg
     1981 gcagagcaat gcctgccagg ccctggacag cctctaccac acccagtgct ttgtttgctg
     2041 ctcttgtggg cgaactttgc gttgcaaggc tttctacagt gtcaatggct ctgtgtactg
     2101 tgaggaagat tatctgtttt cagggtttca ggaggcagct gagaaatgct gtgtctgtgg
     2161 tcacttgatt ttggagaaga tcctacaagc aatggggaag tcctatcatc caggctgttt
     2221 ccgatgcatt gtttgcaaca agtgcctgga tggcatcccc ttcacagtgg acttctccaa
     2281 ccaagtatac tgtgtcaccg actaccacaa aaattatgct cctaagtgtg cagcctgtgg
     2341 ccaacccatc ctcccctctg agggctgtga ggacatcgtg agggtgatat ccatggaccg
     2401 ggattatcac tttgagtgct accactgtga ggactgccgg atgcagctga gtgatgagga
     2461 aggctgctgc tgtttccctc tggatgggca cttgctctgc catggttgcc acatgcagcg
     2521 gctcaatgcc cgacaacccc ctgccaacta tatcctcgag gactacaaag accatgacgg
     2581 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     2641 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2701 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2761 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2821 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2881 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2941 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3001 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3061 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3121 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3181 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3241 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3301 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3361 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3421 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3481 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3541 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3601 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3661 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3721 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3781 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3841 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3901 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3961 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4021 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4081 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4141 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4201 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4261 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4321 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4381 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4441 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4501 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4561 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4621 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4681 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4741 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4801 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4861 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4921 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4981 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5041 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5101 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5161 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5221 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5821 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7021 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
