LOCUS       V036119                 7435 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036119
VERSION     V036119
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             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2955
                     /label="KATNB1(NM_005886)"
                     /note="KATNB1(NM_005886)"
                     /gene="KATNB1"
     polyA_signal    3035..3259
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3305..3733
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3747..4076
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4143..4937
                     /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    5111..5232
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5281..5297)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5305..5321
                     /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(5329..5359)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5374..5395
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5683..6268)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6439..7299)
                     /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(7300..7404)
                     /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 ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggccaccc ctgtggtcac caagacagcc
     1021 tggaagttgc aagagatcgt cgcgcatgcc agcaacgtgt cctcactggt gctgggcaaa
     1081 gcctccgggc ggctgctggc tacaggcggg gatgactgcc gcgtcaacct gtggtccatc
     1141 aacaagccca actgcatcat gagcctgacg ggccacacat ccccagtgga gagcgtccgc
     1201 ctcaacaccc ccgaggagct catcgtggcc ggctctcagt cgggctccat ccgtgtctgg
     1261 gacctggaag ctgccaaaat tcttcgcaca ctcatgggac acaaagccaa catctgcagc
     1321 ctggatttcc acccgtacgg cgagtttgta gcctctggtt cccaggacac aaacatcaag
     1381 ctctgggaca tcaggaggaa aggctgtgtc ttccgataca gggggcacag ccaggccgtg
     1441 cggtgtctcc ggttcagccc cgatgggaag tggttggcgt cggccgcaga tgaccacacc
     1501 gtgaagctct gggatctcac tgccggcaag atgatgtctg agttccctgg tcacacgggg
     1561 cctgtcaacg tggtcgagtt tcaccccaac gagtacctcc tggcctccgg cagctctgac
     1621 aggacaatcc gcttctggga cctggagaag ttccaggtgg tgagctgcat cgaaggggag
     1681 cctgggcccg tcaggagcgt cctcttcaac ccagacggct gctgcctgta cagcggctgc
     1741 caggactcac tgcgtgtcta cggctgggaa cctgagcggt gctttgatgt ggtcctcgtc
     1801 aactggggca aggtggccga cctggccatc tgcaatgacc agttgatagg tgtggccttc
     1861 tcccagagca acgtctcctc ctacgtggtg gatctgacgc gtgtcaccag gactggcacg
     1921 gtggcccggg accctgtgca ggaccaccgg cccctggcac agccactgcc caaccccagc
     1981 gcccccctcc ggcgcatcta tgagcggccc agcacaacct gcagcaagcc tcagagggtg
     2041 aagcagaact cagagagcga gcgccgcagc cccagcagcg aggatgaccg ggacgagcgc
     2101 gagtcccgcg cggagatcca gaacgccgag gactacaacg agatcttcca gcccaagaac
     2161 agcatcagtc ggacgccacc ccggagaagt gagcccttcc ctgcaccccc agaggacgac
     2221 gcagccacag caaaggaggc agcaaagccc agccctgcca tggatgtgca gttcccggtg
     2281 ccaaatctgg aggtcctgcc ccggccccca gtggttgctt ccacacctgc acccaaggct
     2341 gagcctgcca tcatccctgc cacccggaac gagcccatcg ggctgaaggc ctccgacttc
     2401 ctgcccgccg tgaagatccc ccagcaggcc gagctggtgg acgaggatgc catgtcacag
     2461 atccgcaaag gccacgacac catgtgtgtg gtgctcacca gccgccacaa gaacctggac
     2521 actgtgcggg ctgtgtggac catgggcgac atcaagacgt cggtggactc cgctgtggcc
     2581 atcaacgacc tgtcggtggt ggtggacctc ctgaacatcg tcaaccagaa agcctccctg
     2641 tggaagctgg acctgtgcac caccgtcctg ccacagattg agaagcttct gcagagcaag
     2701 tatgagagct acgtccagac gggctgcacc tccctgaagc tgatcctgca gcggtttctg
     2761 cccctcatca cagacatgct ggcggcccca ccctctgtgg gtgtggatat cagcagggag
     2821 gagaggctgc ataagtgccg gctctgctac aagcagctta agagcatcag cggcctggtc
     2881 aagagcaagt caggcctgag cggccgccat ggcagtacct tccgcgagct gcacctgctc
     2941 atggccagtc tggactgaga attctgcaga tatccagcac agtggcggcc gctcgagtct
     3001 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3061 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3121 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3181 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3241 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3301 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3361 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3421 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3481 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3541 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3601 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3661 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3721 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4081 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4141 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4201 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4261 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4321 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4381 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4441 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4501 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4561 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4621 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4681 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4741 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4801 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4861 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4921 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4981 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5041 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5101 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5161 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5221 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5281 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5341 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5401 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5461 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5521 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5581 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5641 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5701 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5761 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5821 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5881 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5941 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6001 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6061 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6121 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6181 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
