LOCUS       V036532                 7628 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036532
VERSION     V036532
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             890..934
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
     CDS             941..1054
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     CDS             1061..1087
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1094..3115
                     /label="ALOX5(NM_000698)"
                     /note="ALOX5(NM_000698)"
                     /gene="ALOX5"
     polyA_signal    3195..3419
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3465..3893
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3907..4236
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4303..5097
                     /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    5271..5392
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5441..5457)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5465..5481
                     /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(5489..5519)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5534..5555
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5843..6428)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6599..7459)
                     /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(7460..7564)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttaccatga aagaaaccgc
      901 tgctgctaaa ttcgaacgcc agcacatgga cagcggtgga atggacgaga agaccaccgg
      961 ctggagaggc ggccacgtgg tggagggcct ggccggcgag ctggagcagc tgagagccag
     1021 actggagcac caccctcagg gccagagaga acctggtacc tacccatacg atgttccaga
     1081 ttacgctgga tccatgccct cctacacggt caccgtggcc actggcagcc agtggttcgc
     1141 cggcactgac gactacatct acctcagcct cgtgggctcg gcgggctgca gcgagaagca
     1201 cctgctggac aagcccttct acaacgactt cgagcgtggc gcggtggatt catacgacgt
     1261 gactgtggac gaggaactgg gcgagatcca gctggtcaga atcgagaagc gcaagtactg
     1321 gctgaatgac gactggtacc tgaagtacat cacgctgaag acgccccacg gggactacat
     1381 cgagttcccc tgctaccgct ggatcaccgg cgatgtcgag gttgtcctga gggatggacg
     1441 cgcaaagttg gcccgagatg accaaattca cattctcaag caacaccgac gtaaagaact
     1501 ggaaacacgg caaaaacaat atcgatggat ggagtggaac cctggcttcc ccttgagcat
     1561 cgatgccaaa tgccacaagg atttaccccg tgatatccag tttgatagtg aaaaaggagt
     1621 ggactttgtt ctgaattact ccaaagcgat ggagaacctg ttcatcaacc gcttcatgca
     1681 catgttccag tcttcttgga atgacttcgc cgactttgag aaaatctttg tcaagatcag
     1741 caacactatt tctgagcggg tcatgaatca ctggcaggaa gacctgatgt ttggctacca
     1801 gttcctgaat ggctgcaacc ctgtgttgat ccggcgctgc acagagctgc ccgagaagct
     1861 cccggtgacc acggagatgg tagagtgcag cctggagcgg cagctcagct tggagcagga
     1921 ggtccagcaa gggaacattt tcatcgtgga ctttgagctg ctggatggca tcgatgccaa
     1981 caaaacagac ccctgcacac tccagttcct ggccgctccc atctgcttgc tgtataagaa
     2041 cctggccaac aagattgtcc ccattgccat ccagctcaac caaatcccgg gagatgagaa
     2101 ccctattttc ctcccttcgg atgcaaaata cgactggctt ttggccaaaa tctgggtgcg
     2161 ttccagtgac ttccacgtcc accagaccat cacccacctt ctgcgaacac atctggtgtc
     2221 tgaggttttt ggcattgcaa tgtaccgcca gctgcctgct gtgcacccca ttttcaagct
     2281 gctggtggca cacgtgagat tcaccattgc aatcaacacc aaggcccgtg agcagctcat
     2341 ctgcgagtgt ggcctctttg acaaggccaa cgccacaggg ggcggtgggc acgtgcagat
     2401 ggtgcagagg gccatgaagg acctgaccta tgcctccctg tgctttcccg aggccatcaa
     2461 ggcccggggc atggagagca aagaagacat cccctactac ttctaccggg acgacgggct
     2521 cctggtgtgg gaagccatca ggacgttcac ggccgaggtg gtagacatct actacgaggg
     2581 cgaccaggtg gtggaggagg acccggagct gcaggacttc gtgaacgatg tctacgtgta
     2641 cggcatgcgg ggccgcaagt cctcaggctt ccccaagtcg gtcaagagcc gggagcagct
     2701 gtcggagtac ctgaccgtgg tgatcttcac cgcctccgcc cagcacgccg cggtcaactt
     2761 cggccagtac gactggtgct cctggatccc caatgcgccc ccaaccatgc gagccccgcc
     2821 accgactgcc aagggcgtgg tgaccattga gcagatcgtg gacacgctgc ccgaccgcgg
     2881 ccgctcctgc tggcatctgg gtgcagtgtg ggcgctgagc cagttccagg aaaacgagct
     2941 gttcctgggc atgtacccag aagagcattt tatcgagaag cctgtgaagg aagccatggc
     3001 ccgattccgc aagaacctcg aggccattgt cagcgtgatt gctgagcgca acaagaagaa
     3061 gcagctgcca tattactact tgtccccaga ccggattccg aacagtgtgg ccatctgaga
     3121 attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg
     3181 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3241 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3301 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3361 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3421 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3481 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3541 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3601 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3661 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3721 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3781 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3841 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4141 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4261 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4321 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4381 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4441 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4501 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4561 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4621 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4681 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4741 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4801 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4861 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4921 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4981 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5041 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5101 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5161 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5221 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5281 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5341 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5401 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5461 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5521 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5581 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5641 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5701 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5761 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5821 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5881 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6181 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6241 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6301 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6361 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6421 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6541 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6661 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6781 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7021 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7261 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7441 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7501 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7561 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7621 cctatggt
//
