LOCUS       V038440                 8325 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038440
VERSION     V038440
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..3005
                     /label="MMUT(NM_000255)"
                     /note="MMUT(NM_000255)"
                     /gene="MMUT"
     CDS             3012..3077
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3101..3689
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3755..3803
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4005..4433
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4447..4776
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4843..5637
                     /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    5813..5934
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5983..5999)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6007..6023
                     /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(6031..6061)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6076..6097
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6385..6973)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7144..8004)
                     /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(8005..8109)
                     /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 ccaccatgtt aagagctaag aatcagcttt
      781 ttttactttc acctcattac ctgaggcagg taaaagaatc atcaggctcc aggctcatac
      841 agcaacgact tctacaccag caacagcccc ttcacccaga atgggctgcc ctggctaaaa
      901 agcagctgaa aggcaaaaac ccagaagacc taatatggca caccccggaa gggatctcta
      961 taaaaccctt gtattccaag agagatacta tggacttacc tgaagaactt ccaggagtga
     1021 agccattcac acgtggacca tatcctacca tgtatacctt taggccctgg accatccgcc
     1081 agtatgctgg ttttagtact gtggaagaaa gcaataagtt ctataaggac aacattaagg
     1141 ctggtcagca gggattatca gttgcctttg atctggcgac acatcgtggc tatgattcag
     1201 acaaccctcg agttcgtggt gatgttggaa tggctggagt tgctattgac actgtggaag
     1261 ataccaaaat tctttttgat ggaattcctt tagaaaaaat gtcagtttcc atgactatga
     1321 atggagcagt tattccagtt cttgcaaatt ttatagtaac tggagaagaa caaggtgtac
     1381 ctaaagagaa gcttactggt accatccaaa atgatatact aaaggaattt atggttcgaa
     1441 atacatacat ttttcctcca gaaccatcca tgaaaattat tgctgacata tttgaatata
     1501 cagcaaagca catgccaaaa tttaattcaa tttcaattag tggataccat atgcaggaag
     1561 caggggctga tgccattctg gagctggcct atactttagc agatggattg gagtactcta
     1621 gaactggact ccaggctggc ctgacaattg atgaatttgc accaaggttg tctttcttct
     1681 ggggaattgg aatgaatttc tatatggaaa tagcaaagat gagagctggt agaagactct
     1741 gggctcactt aatagagaaa atgtttcagc ctaaaaactc aaaatctctt cttctaagag
     1801 cacactgtca gacatctgga tggtcactta ctgagcagga tccctacaat aatattgtcc
     1861 gtactgcaat agaagcaatg gcagcagtat ttggagggac tcagtctttg cacacaaatt
     1921 cttttgatga agctttgggt ttgccaactg tgaaaagtgc tcgaattgcc aggaacacac
     1981 aaatcatcat tcaagaagaa tctgggattc ccaaagtggc tgatccttgg ggaggttctt
     2041 acatgatgga atgtctcaca aatgatgttt atgatgctgc tttaaagctc attaatgaaa
     2101 ttgaagaaat gggtggaatg gccaaagctg tagctgaggg aatacctaaa cttcgaattg
     2161 aagaatgtgc tgcccgaaga caagctagaa tagattctgg ttctgaagta attgttggag
     2221 taaataagta ccagttggaa aaagaagacg ctgtagaagt tctggcaatt gataatactt
     2281 cagtgcgaaa caggcagatt gaaaaactta agaagatcaa atccagcagg gatcaagctt
     2341 tggctgaacg ttgtcttgct gcactaaccg aatgtgctgc tagcggagat ggaaatatcc
     2401 tggctcttgc agtggatgca tctcgggcaa gatgtacagt gggagaaatc acagatgccc
     2461 tgaaaaaggt atttggtgaa cataaagcga atgatcgaat ggtgagtgga gcatatcgcc
     2521 aggaatttgg agaaagtaaa gagataacat ctgctatcaa gagggttcat aaattcatgg
     2581 aacgtgaagg tcgcagacct cgtcttcttg tagcaaaaat gggacaagat ggccatgaca
     2641 gaggagcaaa agttattgct acaggatttg ctgatcttgg ttttgatgtg gacataggcc
     2701 ctcttttcca gactcctcgt gaagtggccc agcaggctgt ggatgcggat gtgcatgctg
     2761 tgggcataag caccctcgct gctggtcata aaaccctagt tcctgaactc atcaaagaac
     2821 ttaactccct tggacggcca gatattcttg tcatgtgtgg aggggtgata ccacctcagg
     2881 attatgaatt tctgtttgaa gttggtgttt ccaatgtatt tggtcctggg actcgaattc
     2941 caaaggctgc cgttcaggtg cttgatgata ttgagaagtg tttggaaaag aagcagcaat
     3001 ctgtaggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3061 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3121 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3181 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3241 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3301 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3361 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3421 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3481 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3541 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3601 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3661 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3721 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3781 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3841 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3901 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3961 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4021 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4081 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4141 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4201 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4261 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4321 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4381 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4441 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4501 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4561 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4621 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4681 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4741 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4801 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4861 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4921 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4981 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5041 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5101 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5161 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5221 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5281 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5341 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5401 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5461 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5521 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5581 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5641 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5701 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5761 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5821 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5881 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5941 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6001 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6061 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6121 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6181 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7021 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7081 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7141 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7201 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7261 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7321 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7381 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7441 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7501 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7561 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7621 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7681 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7741 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7801 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7861 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7921 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7981 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8041 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8101 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8161 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8221 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8281 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
