LOCUS       V040111                 7255 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040111
VERSION     V040111
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1063..2775
                     /label="KLHL28(NM_017658)"
                     /note="KLHL28(NM_017658)"
                     /gene="KLHL28"
     polyA_signal    2855..3079
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3125..3553
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3567..3896
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3963..4757
                     /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    4931..5052
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5101..5117)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5125..5141
                     /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(5149..5179)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5194..5215
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5503..6088)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6259..7119)
                     /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(7120..7224)
                     /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 aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttggcggcgg agggagtgga ggagggggaa gcatggacca cacatccccg
     1081 acctacatgc ttgctaactt aacccacttg cattctgaac aacttctgca gggcttgaat
     1141 cttcttcgcc aacatcacga actctgtgac atcattcttc gagtaggtga tgttaaaatt
     1201 catgctcaca aagtggtact tgccagcgtc agcccgtatt tcaaagctat gttcactgga
     1261 aacctttctg aaaaagagaa cagtgaggtt gagtttcaat gcattgatga aactgctctc
     1321 caggccattg tggagtatgc ctatacaggg actgttttta tttctcagga cacagttgaa
     1381 tctctcctgc cagcagcaaa cctactccag ataaaacttg tcctgaaaga atgttgtgca
     1441 tttcttgaaa gccaacttga tcctggtaat tgtattggaa tttctcgttt tgcagaaaca
     1501 tatggttgcc gtgaccttta tttggcagcc actaaataca tatgccagaa ttttgaagct
     1561 gtttgccaga ctgaagagtt ttttgagctt acacatgctg acttggatga aattgtttcc
     1621 aatgactgtt tgaatgtagc taccgaagag actgtttttt atgcattaga gtcttggatc
     1681 aagtatgatg tacaagaacg ccagaaatac ttagcacagt tactaaacag tgtacgatta
     1741 ccattgttga gtgttaagtt tctcactaga ctatatgaag caaatcatct tattcgtgat
     1801 gatcgcactt gtaaacatct tttgaatgaa gccctaaagt accactttat gcctgaacat
     1861 agactctctc atcagacagt cttgatgaca cgacctcgct gtgctcccaa agtactttgt
     1921 gcagtaggag ggaaatctgg actctttgcc tgtttggata gtgtggagat gtactttcct
     1981 cagaatgact cttggattgg tttggcaccc ctaaacattc ctcgctatga atttggaata
     2041 tgcgttttag accaaaaagt atatgttata ggtggtattg caactaatgt gcgtcctggc
     2101 gtcactatca gaaaacatga aaattcagtg gaatgctgga atcctgatac aaatacttgg
     2161 acttctctag agagaatgaa tgaaagccga agtactcttg gagtagtagt acttgcagga
     2221 gaactttatg ccttaggtgg ttatgatgga caatcttatt tacaatctgt agagaagtac
     2281 attcccaaaa taagaaaatg gcaacctgtg gcaccaatga cgacaacaag aagttgtttt
     2341 gctgcagcgg tattggatgg aatgatatat gccattggtg ggtatggtcc tgcccacatg
     2401 aacagtgtgg agcgttatga tccaagtaag gactcctggg agatggttgc atccatggca
     2461 gataaaagga ttcactttgg cgtgggtgtc atgctaggct ttatttttgt ggtgggtgga
     2521 cataatggag tctcacattt gtccagcatt gaaagatacg atcctcatca aaatcagtgg
     2581 actgtgtgta gaccaatgaa agaacctaga acaggagttg gtgctgctgt aatcgataac
     2641 tacctttatg tcgtcggtgg tcactcaggg tcttcctatc tgaatacagt gcagaaatat
     2701 gatcctatct cagatacgtg gctggattca gctggcatga tatactgtcg ctgcaacttt
     2761 gggttaactg cactttgaga attctgcaga tatccagcac agtggcggcc gctcgagtct
     2821 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2881 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2941 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3001 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3061 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3121 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3181 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3241 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3301 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3361 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3421 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3481 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3541 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3901 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4141 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4741 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4801 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4861 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4921 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4981 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5041 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5101 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5161 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5221 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5281 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5341 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5401 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5461 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5521 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5581 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5641 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5701 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5761 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5821 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5881 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5941 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6001 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6061 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6121 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6181 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6241 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6301 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6361 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6421 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6481 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6541 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6601 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6661 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6721 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6781 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6841 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6901 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6961 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7021 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7081 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7141 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7201 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
