LOCUS       V045076                 8287 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045076
VERSION     V045076
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..3777
                     /label="Cblb(NM_001033238)"
                     /note="Cblb(NM_001033238)"
                     /gene="Cblb"
     CDS             3784..3849
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3887..4111
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4157..4585
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4599..4928
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4995..5789
                     /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    5963..6084
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6133..6149)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6157..6173
                     /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(6181..6211)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6226..6247
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6535..7120)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7291..8151)
                     /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(8152..8256)
                     /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 accatggcaa attctatgaa tggcagaaat cctggtggtc gaggaggaaa cccccgcaaa
     1021 ggtcgcattt tggggattat tgatgctatt caggatgcag tcggaccccc aaagcaagct
     1081 gcagctgacc gcaggactgt ggagaagact tggaaactca tggacaaagt ggtaagactg
     1141 tgccaaaatc ccaagcttca gttgaaaaac agcccaccgt atatacttga tattttacct
     1201 gatacgtatc agcacttgag acttatattg agtaaatatg atgacaacca gaagctggct
     1261 caactgagcg agaatgagta ctttaaaatc tacatcgata gtctcatgaa gaagtcgaag
     1321 cgagcgatcc ggctctttaa agaaggcaag gaaaggatgt acgaagagca gtcgcaggac
     1381 agacggaatc tcacaaagct gtcccttatc ttcagtcaca tgctggcaga aatcaaggcg
     1441 atctttccca atggccagtt ccagggagat aacttccgga tcaccaaagc agatgctgct
     1501 gagttctgga ggaagttttt tggagacaaa actattgtac catggaaagt cttcagacag
     1561 tgcctgcatg aggtccatca gatcagctct ggcctggaag caatggctct gaagtcaacc
     1621 attgatttaa cttgcaatga ttacatctca gtgtttgaat ttgatatttt taccaggcta
     1681 tttcagccct ggggctctat tttacggaat tggaacttct tggctgtgac tcacccggga
     1741 tacatggcat ttctcacgta tgatgaagtt aaagctcggc tacagaaata cagcaccaag
     1801 cctggaagtt acattttccg gttaagctgc actcggctgg gacaatgggc cattggctat
     1861 gtgactgggg acggcaatat cctacagacc atacctcata acaagcccct gttccaagcc
     1921 ctgattgatg gtagcaggga aggcttttat ctttatcctg atgggcggag ctataaccct
     1981 gatttaaccg gattatgtga acctacacct cacgatcata taaaagttac acaggagcag
     2041 tatgaactgt attgtgaaat gggctccact tttcagctct gcaagatctg tgcagagaat
     2101 gacaaagatg tcaagattga gccttgcggg catctgatgt gcacttcgtg ccttaccgcg
     2161 tggcaggagt ctgatggcca aggctgcccc ttctgtcgct gtgagataaa aggaacggag
     2221 cccatcattg tggacccctt cgaccccaga gatgaaggct ccaggtgctg cagcatcatt
     2281 gaccctttca gcatccccat gcttgacttg gacgatgacg atgatcgaga ggagtctttg
     2341 atgatgaaca ggctggcgag tgttcgaaag tgcacggaca ggcagaactc accagtcaca
     2401 tcgccaggct cctcacccct tgcccagaga agaaagcctc agccagaccc tctccagatc
     2461 ccccacctca gcctgccacc agtgcctccc cgcctagatc tcattcagaa aggcatcgtg
     2521 cgctctccgt gtggcagccc cacaggctcg ccaaagtctt ctccatgcat ggttagaaaa
     2581 caagacaaac cactcccggc accccctcct cccttgagag atccacctcc tccaccagag
     2641 aggcctcccc ccatcccacc tgacaataga ctgagcagac acttccacca tggagagagt
     2701 gtgccttccc gggaccagcc catgcccctc gaagcctggt gccctcggga tgccttcggg
     2761 actaaccagg tgatgggatg tcgcatcctc ggggatggct ctccaaagcc tggcgtcacc
     2821 gcaaactcca gcttaaatgg gaggcacagt agaatgggct ccgagcaggt tcttatgagg
     2881 aaacacagac gccatgattt gccttcagaa ggagccaagg tcttttccaa tggccacctt
     2941 gccactgaag aatacgatgt ccctcctcgg ctttcccctc cgcctccagt caccaccctt
     3001 ctcccaagca taaagtgtac tggtccatta gcaaattgtc tctcagagaa aactagagac
     3061 acggtagaag atgatgacga tgaatacaag attccttcat cccatcctgt gtccctgaat
     3121 tcacaaccat ctcattgcca taatgtcaaa gctcctgttc ggtcttgtga taatggtcat
     3181 tgtatattga atggaactca tggtgcgcct tcagagatga agaaatcgaa catcccagat
     3241 ttaggcatct atttgaaggg tgaggacgcc tttgatgctc tccctccatc cctccctcct
     3301 cccccacctc ctgcaagaca cagtctcatc gaacattcaa aacctccagg ctccagtagc
     3361 cggccttcct cagggcagga ccttttcctt cttccttcag atcccttttt tgacccaaca
     3421 agtggccaag ttccattgcc tcctgccagg agagcagcag gagacagcgg caaagccaac
     3481 agagcctcgc aggactatga ccagctccct tcatcttccg atggttcgca agcaccagct
     3541 agacccccca aaccacgacc ccgaaggact gcaccagaaa ttcatcacag aaagccccat
     3601 gggcccgaag cggcgttgga aaatgtcgat gcaaaaattg caaaactcat gggagagggg
     3661 tatgcctttg aagaggtgaa gagagcctta gagattgccc agaataacgt ggaagtggcc
     3721 aggagcatac ttcgagaatt tgccttcccc cctcctgtct ccccacgtct gaatctactc
     3781 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3841 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3901 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3961 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4021 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4081 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4141 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4201 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4261 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4321 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4381 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4441 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4501 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4561 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4621 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4681 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4741 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4801 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4861 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4921 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4981 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5041 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5101 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5161 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5221 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5281 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5341 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5401 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5461 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5521 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5581 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5641 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5701 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5761 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5821 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5881 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5941 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6061 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6121 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     7081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8281 tgacgtc
//
