LOCUS       V026273                 8208 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026273
VERSION     V026273
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2888
                     /label="ALOXE3(NM_021628)"
                     /note="ALOXE3(NM_021628)"
                     /gene="ALOXE3"
     CDS             2895..2960
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2984..3572
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3638..3686
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3888..4316
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4330..4659
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4726..5520
                     /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    5696..5817
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5866..5882)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5890..5906
                     /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(5914..5944)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5959..5980
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6268..6856)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7027..7887)
                     /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(7888..7992)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc agtgtaccgc ctgtgtgtga
      781 ccactggtcc ctacctgagg gccggcacac tggacaacat ctctgtcaca ctggtgggca
      841 cgtgtggtga aagccccaag cagcggctag atcgaatggg cagggacttc gcccctggat
      901 cggtacagaa gtacaaggtg cgttgcacag cggagctggg tgagctcttg ctgctgcgtg
      961 tacacaagga gcgctacgct ttcttccgca aggactcttg gtactgtagc cgcatctgtg
     1021 tcaccgaacc ggatggtagt gtatcccact tcccctgcta tcagtggatt gaaggctact
     1081 gcaccgtgga gctgaggcca ggaacagcaa gaactatttg tcaggactct cttcccctcc
     1141 tcctggatca caggacacgg gagctccggg cccgacaaga atgctaccgc tggaagatct
     1201 atgcccctgg cttcccctgc atggtagacg tcaacagctt tcaggagatg gagtcagaca
     1261 agaaatttgc cttgacaaag acgacaactt gtgtagacca gggtgacagc agtgggaatc
     1321 ggtacctgcc cggcttcccc atgaaaattg acatcccatc cctgatgtac atggagccca
     1381 atgttcgata ctcagccacc aagacgatct cgctgctctt caatgccatc cctgcgtcct
     1441 tgggaatgaa gcttcgaggg ctgttggatc gcaagggctc ctggaagaag ctggatgaca
     1501 tgcagaacat cttctggtgc cataagacct tcacgacaaa gtatgtcaca gagcactggt
     1561 gtgaagatca cttctttggg taccagtacc tgaatggtgt caatcccgtc atgctccact
     1621 gcatctctag cttgcccagc aagctgcctg tcaccaatga catggtggcc cccttgctgg
     1681 gacaggacac atgcctgcag acagagctag agagggggaa catcttccta gcggactact
     1741 ggatcctggc ggaggccccc acccactgcc taaacggccg ccagcagtac gtggccgccc
     1801 cactgtgcct gctgtggctc agcccccagg gggcgctggt gcccttggcc atccagctca
     1861 gccagacccc cgggcctgac agccccatct tcctgcccac tgactccgaa tgggactggc
     1921 tgctggccaa gacgtgggtg cgcaactctg agttcctggt gcacgaaaac aacacgcact
     1981 ttctgtgcac gcatttgctg tgcgaggcct tcgccatggc cacgctgcgc cagctgccgc
     2041 tctgccaccc catctacaag ctcctactcc cccacactcg atacacgctg caggtgaaca
     2101 ccatcgcgag ggccacgctg ctcaaccccg agggcctcgt ggaccaggtc acgtccatcg
     2161 ggaggcaagg cctcatctac ctcatgagca cgggcctggc ccacttcacc tacaccaatt
     2221 tctgccttcc ggacagcctg cgggcccgcg gcgtcctggc tatccccaac taccactacc
     2281 gagacgacgg cctgaagatc tgggcggcca ttgagagctt tgtctcagaa atcgtgggct
     2341 actattatcc cagtgacgca tctgtgcagc aggattcgga gctgcaggcc tggactggcg
     2401 agatttttgc tcaggcgttc ctgggccggg aaagctcagg tttcccaagc cggctgtgca
     2461 ccccaggaga gatggtgaag ttcctcactg caatcatctt caattgctct gcccagcacg
     2521 ctgctgtcaa cagtgggcag catgactttg gggcctggat gcccaatgct ccatcatcca
     2581 tgaggcagcc cccaccccag accaagggga ccaccaccct gaagacttac ctagacaccc
     2641 tccctgaagt gaacatcagc tgtaacaacc tcctcctctt ctggttggtt agccaagaac
     2701 ccaaggacca gaggcccctg ggcacctacc cagatgagca cttcacagag gaggccccga
     2761 ggcggagcat cgccgccttc cagagccgcc tggcccagat ctcaagggac atccaggagc
     2821 ggaaccaggg tctggcactg ccctacacct acctggaccc tcccctcatt gagaacagcg
     2881 tctccatcgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2941 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     3001 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3061 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3121 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3181 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3241 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3301 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3361 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     3421 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3481 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3541 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3601 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3661 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3721 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3781 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3841 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3901 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3961 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4021 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     4081 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4141 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4201 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     4261 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4321 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4381 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4501 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4561 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4621 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4681 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4741 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4801 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4861 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4921 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4981 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5041 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5101 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5161 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5221 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5281 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5341 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5401 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5461 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5521 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5581 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5641 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5701 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5761 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5821 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5881 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5941 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6001 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6061 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6121 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6181 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6241 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6301 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6361 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6421 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6481 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6541 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6601 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6661 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6721 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6781 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6841 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6901 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6961 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7021 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7081 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7141 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7201 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7261 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7321 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7381 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7441 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7501 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7561 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7621 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7681 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7741 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7801 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7861 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7921 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7981 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     8041 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     8101 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     8161 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
