LOCUS       V033829                 7240 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033829
VERSION     V033829
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"
     CDS             964..2730
                     /label="Grn(NM_008175)"
                     /note="Grn(NM_008175)"
                     /gene="Grn"
     CDS             2737..2802
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2840..3064
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3110..3538
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3552..3881
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3948..4742
                     /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    4916..5037
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5086..5102)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5110..5126
                     /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(5134..5164)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5179..5200
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5488..6073)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6244..7104)
                     /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(7105..7209)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgtggg tcctgatgag ctggctggcc ttcgcggcag ggctggtagc cggaacacag
     1021 tgtccagatg ggcagttctg ccctgttgcc tgctgccttg accagggagg agccaactac
     1081 agctgctgta accctcttct ggacacatgg cctagaataa cgagccatca tctagatggc
     1141 tcctgccaga cccatggcca ctgtcctgct ggctattctt gtcttctcac tgtgtctggg
     1201 acttccagct gctgcccgtt ctctaagggt gtgtcttgtg gtgatggcta ccactgctgc
     1261 ccccagggct tccactgtag tgcagatggg aaatcctgct tccagatgtc agataacccc
     1321 ttgggtgctg tccagtgtcc tgggagccag tttgaatgtc ctgactctgc cacctgctgc
     1381 attatggttg atggttcgtg gggatgttgt cccatgcccc aggcctcttg ctgtgaagac
     1441 agagtgcatt gctgtcccca tggggcctcc tgtgacctgg ttcacacacg atgcgtttca
     1501 cccacgggca cccacaccct actaaagaag ttccctgcac aaaagaccaa cagggcagtg
     1561 tctttgcctt tttctgtcgt gtgccctgat gctaagaccc agtgtcccga tgattctacc
     1621 tgctgtgagc tacccactgg gaagtatggc tgctgtccaa tgcccaatgc catctgctgt
     1681 tccgaccacc tgcactgctg cccccaggac actgtatgtg acctgatcca gagtaagtgc
     1741 ctatccaaga actacaccac ggatctcctg accaagctgc ctggataccc agtgaaggag
     1801 gtgaagtgcg acatggaggt gagctgccct gaaggatata cctgctgccg cctcaacact
     1861 ggggcctggg gctgctgtcc atttgccaag gccgtgtgtt gtgaggatca cattcattgc
     1921 tgcccggcag ggtttcagtg tcacacagag aaaggaacct gcgaaatggg tatcctccaa
     1981 gtaccctgga tgaagaaggt catagccccc ctccgcctgc cagacccaca gatcttgaag
     2041 agtgatacac cttgtgatga cttcactagg tgtcctacaa acaatacctg ctgcaaactc
     2101 aattctgggg actggggctg ctgtcccatc ccagaggctg tctgctgctc agacaaccag
     2161 cattgctgcc ctcagggctt cacatgtctg gctcaggggt actgtcagaa gggagacaca
     2221 atggtggctg gcctggagaa gatacctgcc cgccagacaa ccccgctcca aattggagat
     2281 atcggttgtg accagcatac cagctgccca gtagggcaaa cctgctgccc aagcctcaag
     2341 ggaagttggg cctgctgcca gctgccccat gctgtgtgct gtgaggaccg gcagcactgt
     2401 tgcccggccg ggtacacctg caatgtgaag gcgaggacct gtgagaagga tgtcgatttt
     2461 atccagcctc ccgtgctcct gaccctcggc cctaaggttg ggaatgtgga gtgtggagaa
     2521 gggcatttct gccatgataa ccagacctgt tgtaaagaca gtgcaggagt ctgggcctgc
     2581 tgtccctacc taaagggtgt ctgctgtaga gatggacgtc actgttgccc cggtggcttc
     2641 cactgttcag ccaggggaac caagtgtttg cgaaagaaga ttcctcgctg ggacatgttt
     2701 ttgagggatc cggtcccaag accgctactg ctcgaggact acaaagacca tgacggtgat
     2761 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgtttaa
     2821 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2881 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2941 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3001 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3061 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3121 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3181 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3241 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3301 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3361 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3421 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3481 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3541 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3781 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3841 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3901 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3961 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4021 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4081 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4141 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4201 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4261 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4321 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4381 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4441 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4501 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4561 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4621 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4681 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4741 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4801 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4861 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4921 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4981 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5041 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5101 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5161 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5221 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5281 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5341 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5401 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5461 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5521 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5581 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5641 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5701 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5761 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5821 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5881 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5941 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6001 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6061 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6121 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6181 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6241 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6301 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6361 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6421 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6481 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6541 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6601 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6661 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6721 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6781 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6841 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6901 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6961 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7021 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7081 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7141 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7201 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
