LOCUS       V032272                 8058 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032272
VERSION     V032272
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..2738
                     /label="Acox1(NM_015729)"
                     /note="Acox1(NM_015729)"
                     /gene="Acox1"
     CDS             2745..2810
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2834..3422
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3488..3536
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3738..4166
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4180..4509
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4576..5370
                     /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    5546..5667
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5716..5732)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5740..5756
                     /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(5764..5794)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5809..5830
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6118..6706)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6877..7737)
                     /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(7738..7842)
                     /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 ccaccatgaa tcccgatctg cgcaaggagc
      781 gggccgccgc caccttcaat ccagagttaa tcacgcacat cttggatggt agtccggaga
      841 acacccggcg ccgtcgagaa atcgagaact tgattctgaa cgatccagac ttccaacatg
      901 aggactataa cttcctcact cgaagccagc gttacgaggt ggctgttaag aagagtgcca
      961 ccatggtgaa gaagatgagg gaatttggca tcgcagaccc tgaagaaatc atgtggttta
     1021 aaaactctgt gcaccgaggg catcctgagc ctttggacct tcacttgggc atgttcctgc
     1081 ccaccttgct tcaccaggcc accgaagagc agcaggagcg tttcttcatg ccggcctgga
     1141 atctggagat cacgggcact tatgcgcaga cagagatggg tcatggaact catcttcgag
     1201 gcttggaaac cactgccaca tatgacccca agacccaaga gttcattctc aacagcccaa
     1261 ctgtgacttc catcaagtgg tggcctgggg ggcttgggaa gacttccaat catgcgatag
     1321 tcctggctca gctcatcact cgaggggagt gctacgggtt acatgccttt gttgtcccta
     1381 tccgtgagat tgggacccac aagcctctgc caggcatcac tgttggggat atcggcccca
     1441 agtttggtta tgaagagatg gataatggct acctgaagat ggacaattac cgtattccta
     1501 gagagaacat gttgatgaaa tatgcccagg tgaagcctga cggcacgtat gtaaaacctc
     1561 tgagtaacaa gctgacatat gggaccatgg ttttcgtaag gtccttcctc gtgggaagtg
     1621 cagctcagag tctgtccaag gcatgcacca ttgccattcg atacagtgct gtgaggcgcc
     1681 agtctgaaat caagagaagc gagccagagc cccagatttt ggattttcag acgcagcagt
     1741 ataaactctt cccgctcctg gccaccgcct atgccttcca ctttctcgga agatacataa
     1801 aggagaccta catgcggatt aatgagagca ttggccaagg cgacctgagt gagctgcctg
     1861 agcttcatgc cctcacagct gggctgaagg cttttactac ctggacagcc aatgctggta
     1921 tcgaagaatg tcggatggct tgcggtgggc acggctattc tcacagcagt gggattccaa
     1981 atatttacgt cacgtttacc ccggcctgca ccttcgaggg ggagaacact gttatgatgc
     2041 tgcagacggc caggttcttg atgaaaatct atgaccaggt tcagtcgggg aagctggtgg
     2101 gtggtatggt gtcgtacttg aatgacctgc cgagccagcg tatccagccg cagcaggtgg
     2161 cagtctggcc aactctggtg gacattaaca gcctggacag cctgacagaa gcctacaagc
     2221 tacgtgcagc cagattggta gaaattgctg caaaaaacct tcaggcccaa gtgagtcaca
     2281 ggaagagcaa ggaagtggcg tggaacttga cttctgtcga ccttgttcgc gcaagtgagg
     2341 cgcactgcca ctacgtgacc gttaaggtct ttgcagataa actccccaag attcaagaca
     2401 gagccgtgca agccgtgctg aggaacctgt gtctcttata ttctctctat gggatcagcc
     2461 agaaaggagg ggattttctt gaggggaaca tcatcacagg ggctcagatg tcacaggtaa
     2521 acagtcggat cctggagctg ctcacagtga ctcgccccaa cgctgtggct ttggtggatg
     2581 cctttgactt taaggatgtg acccttggct ctgttctcgg ccgctatgat ggcaatgtgt
     2641 atgaaaactt gtttgagtgg gccaagaagt ccccactgaa caagacagag gtccacgaat
     2701 cttactacaa gcacttgaag cccctgcagt cgaagcttgg atccgactac aaagaccatg
     2761 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2821 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2881 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2941 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3001 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3061 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3121 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3181 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3241 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3301 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3361 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3421 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3481 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3541 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3601 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3661 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     3721 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3781 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3841 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3901 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3961 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4021 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4081 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     4141 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4261 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4321 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4381 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4441 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4501 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4561 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4621 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4681 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4741 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4801 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4861 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4921 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4981 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5041 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5101 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5161 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5221 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5281 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5341 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     5401 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5461 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5521 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5641 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5701 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5761 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5821 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5881 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5941 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6001 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6061 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6121 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6181 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6241 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6301 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6361 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6421 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6481 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6541 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6601 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6661 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6721 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6781 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6841 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6901 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6961 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7021 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7081 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7141 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7201 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7261 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7321 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7381 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7441 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7501 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7561 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7621 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7681 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7741 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7801 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7861 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7921 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7981 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8041 atgaagaatc tgcttagg
//
