LOCUS       V029300                 8285 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029300
VERSION     V029300
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..3775
                     /label="UNC5B(NM_170744)"
                     /note="UNC5B(NM_170744)"
                     /gene="UNC5B"
     CDS             3782..3847
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3885..4109
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4155..4583
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4597..4926
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4993..5787
                     /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    5961..6082
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6131..6147)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6155..6171
                     /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(6179..6209)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6224..6245
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6533..7118)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7289..8149)
                     /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(8150..8254)
                     /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 atgggggccc ggagcggagc
      961 tcggggcgcg ctgctgctgg cactgctgct ctgctgggac ccgaggctga gccaagcagg
     1021 cactgattct ggcagcgagg tgctccctga ctccttcccg tcagcgccag cagagccgct
     1081 gccctacttc ctgcaggagc cacaggacgc ctacattgtg aagaacaagc ctgtggagct
     1141 ccgctgccgc gccttccccg ccacacagat ctacttcaag tgcaacggcg agtgggtcag
     1201 ccagaacgac cacgtcacac aggaaggcct ggatgaggcc accggcctgc gggtgcgcga
     1261 ggtgcagatc gaggtgtcgc ggcagcaggt ggaggagctc tttgggctgg aggattactg
     1321 gtgccagtgc gtggcctgga gctccgcggg caccaccaag agtcgccgag cctacgtccg
     1381 catcgcctac ctgcgcaaga acttcgatca ggagcctctg ggcaaggagg tgcccctgga
     1441 ccatgaggtt ctcctgcagt gccgcccgcc ggagggggtg cctgtggccg aggtggaatg
     1501 gctcaagaat gaggatgtca tcgaccccac ccaggacacc aacttcctgc tcaccatcga
     1561 ccacaacctc atcatccgcc aggcccgcct gtcggacact gccaactata cctgcgtggc
     1621 caagaacatc gtggccaaac gccggagcac cactgccacc gtcatcgtct acgtgaatgg
     1681 cggctggtcc agctgggcag agtggtcacc ctgctccaac cgctgtggcc gaggctggca
     1741 gaagcgcacc cggacctgca ccaaccccgc tccactcaac ggaggggcct tctgcgaggg
     1801 ccaggcattc cagaagaccg cctgcaccac catctgccca gtcgatgggg cgtggacgga
     1861 gtggagcaag tggtcagcct gcagcactga gtgtgcccac tggcgtagcc gcgagtgcat
     1921 ggcgccccca ccccagaacg gaggccgtga ctgcagcggg acgctgctcg actctaagaa
     1981 ctgcacagat gggctgtgca tgcaaaataa gaaaactcta agcgacccca acagccacct
     2041 gctggaggcc tcaggggatg cggcgctgta tgcggggctc gtggtggcca tcttcgtggt
     2101 cgtggcaatc ctcatggcgg tgggggtggt ggtgtaccgc cgcaactgcc gtgacttcga
     2161 cacagacatc actgactcat ctgctgccct gactggtggt ttccaccccg tcaactttaa
     2221 gacggcaagg cccagcaacc cgcagctcct acacccctct gtgcctcctg acctgacagc
     2281 cagcgccggc atctaccgcg gacccgtgta tgccctgcag gactccaccg acaaaatccc
     2341 catgaccaac tctcctctgc tggacccctt acccagcctt aaggtcaagg tctacagctc
     2401 cagcaccacg ggctctgggc caggcctggc agatggggct gacctgctgg gggtcttgcc
     2461 gcctggcaca taccctagcg atttcgcccg ggacacccac ttcctgcacc tgcgcagcgc
     2521 cagcctcggt tcccagcagc tcttgggcct gccccgagac ccagggagca gcgtcagcgg
     2581 cacctttggc tgcctgggtg ggaggctcag catccccggc acaggggtca gcttgctggt
     2641 gcccaatgga gccattcccc agggcaagtt ctacgagatg tatctactca tcaacaaggc
     2701 agaaagtacc ctcccgcttt cagaagggac ccagacagta ttgagcccct cggtgacctg
     2761 tggacccaca ggcctcctgc tgtgccgccc cgtcatcctc accatgcccc actgtgccga
     2821 agtcagtgcc cgtgactgga tctttcagct caagacccag gcccaccagg gccactggga
     2881 ggaggtggtg accctggatg aggagaccct gaacacaccc tgctactgcc agctggagcc
     2941 cagggcctgt cacatcctgc tggaccagct gggcacctac gtgttcacgg gcgagtccta
     3001 ttcccgctca gcagtcaagc ggctccagct ggccgtcttc gcccccgccc tctgcacctc
     3061 cctggagtac agcctccggg tctactgcct ggaggacacg cctgtagcac tgaaggaggt
     3121 gctggagctg gagcggactc tgggcggata cttggtggag gagccgaaac cgctaatgtt
     3181 caaggacagt taccacaacc tgcgcctctc cctccatgac ctcccccatg cccattggag
     3241 gagcaagctg ctggccaaat accaggagat ccccttctat cacatttgga gtggcagcca
     3301 gaaggccctc cactgcactt tcaccctgga gaggcacagc ttggcctcca cagagctcac
     3361 ctgcaagatc tgcgtgcggc aagtggaagg ggagggccag atattccagc tgcataccac
     3421 tctggcagag acacctgctg gctccctgga cactctctgc tctgcccctg gcagcactgt
     3481 caccacccag ctgggacctt atgccttcaa gatcccactg tccatccgcc agaagatatg
     3541 caacagccta gatgccccca actcacgggg caatgactgg cggatgttag cacagaagct
     3601 ctctatggac cggtacctga attactttgc caccaaagcg agccccacgg gtgtgatcct
     3661 ggacctctgg gaagctctgc agcaggacga tggggacctc aacagcctgg cgagtgcctt
     3721 ggaggagatg ggcaagagtg agatgctggt ggctgtggcc accgacgggg actgcctcga
     3781 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3841 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3901 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3961 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4021 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4081 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4141 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4201 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4261 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4321 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4381 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4441 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4501 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4561 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4621 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4681 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4741 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4801 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4861 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4921 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4981 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5161 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5521 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5761 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5821 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5941 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6001 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6061 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6121 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6181 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6241 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6961 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     7081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8281 acgtc
//
