LOCUS       V043129                 7999 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043129
VERSION     V043129
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..3489
                     /label="Gpld1(NM_008156)"
                     /note="Gpld1(NM_008156)"
                     /gene="Gpld1"
     CDS             3496..3561
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3599..3823
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3869..4297
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4311..4640
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4707..5501
                     /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    5675..5796
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5845..5861)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5869..5885
                     /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(5893..5923)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5938..5959
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6247..6832)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7003..7863)
                     /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(7864..7968)
                     /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 accatgtctg caggcaggct gtggtctagc ctgctgcttc tgctgcctct tttctgctct
     1021 aaaagctcat cttgtggtct ctcaacacat gtagaaatag gacacagggc tctggagttt
     1081 cttcggcttc aagatggacg cattaactac aaagagctga tcttagagca ccaggacgca
     1141 tatcaggctg ggaccgtgtt tcctgatgcc ttttatccta gcatctgcaa aagaggaaaa
     1201 tatcatgacg tttctgagag gactcactgg actccatttc ttaacgccag catccattat
     1261 attcgagaga actaccctct gccctgggag aaggacacag agaagttggt ggctttcttg
     1321 tttggaatca cctcccacat ggtcgctgac gtgagctggc atagcctggg tattgaacaa
     1381 gggttcctca ggacaatggg agctatcgat ttttacaact cttactctga cgctcactcg
     1441 gctggtgatt ttggaggaga tgtgttgagc cagtttgaat ttaattttaa ttacctctca
     1501 cggcgctggt acgtgcccgt cagggatctt ctgagaattt atgataatct ctatggtcgg
     1561 aaagtcatca ccaaagacgt ccttgttgat tgcacctacc ttcagttcct ggaaatgcac
     1621 ggggagatgt ttgctgtttc caagctctat tccacgtact ctacaaagtc cccatttctg
     1681 gtggagcagt tccaagacta tttcctcgga ggtctggatg acatggcatt ctggtccacg
     1741 aacatttacc gtttgaccag ctttatgctg gagaacggga ccagtgactg caacctgcct
     1801 gagaaccccc tgttcatctc ctgtgatggc aggaaccaca ccctcagtgg ctcaaaagtg
     1861 cagaaaaatg attttcacag gaatttgacc atgttcataa gtagagacat caggaaaaac
     1921 ctcaattaca cagaaagagg cgtgttctac agcacaggct cctgggcccc ggaatctgtc
     1981 acctttatgt accagactct ggagaggaac ctgaggctga tgcttgctgg cagctctcag
     2041 aaaaatctga atcatgtctc cagcccctct gcctcctaca ccctgtctgt cccctatgcc
     2101 aggctcggct gggtcatgac ctcagctgac ctcaaccagg atgggcacgg agacctcgtg
     2161 gtgggtgcac caggctacag ccaccctggc cgcttccaga ttgggcgggt gtacatcatc
     2221 tatggcaacg atctgggcct gcctcccatc gacctggact tagacaagga ggcccatggg
     2281 atcctggagg gcttccagcc ttcaggtcgc tttggctcag ctttggccgt gctggacttc
     2341 aaccaggatg ggctgcctga cctggctgtg ggcgctccct ccgtgggctc agggcagctc
     2401 acctacaatg gctcggtgta tgtctactat ggttcccagc aagggaggct gtcttcttcc
     2461 cctaacgtca ccatctcctg caaggacacc tactgtaact tgggctggac cctcttggca
     2521 acagatgcag acggagatgg ccggcacgat ctggtcatca gctccccttt cgcaccaggt
     2581 ggagggaagc agagaggaat tgtggccacc ttttattctc accccagacg gaatgacaaa
     2641 gaattactga ctttggagga agctgactgg aaggtgaatg gggaggagga cttctcgtgg
     2701 tttggatact cccttcacgg ggtcacagtg gcaaacagaa gcctgctgct gattgggagc
     2761 ccgacctgga agaatgtcag caggatggca cgctcaagcc acaaaaagaa ccaagaggaa
     2821 aagagcctgg gaaaggtgta cggctacttt ctcccgaacc gccagagcac aatcactatc
     2881 tcgggagata aggcgatggg aaagctgggt acctccctgt ccagtggtta tgtgagagtg
     2941 aatgggaccc taacccaagt cctgctggtt ggagccccta ctcatgatga cgtgtctaaa
     3001 atggcattcc taaccatgac cctgcaccag ggtggagcca ctcggatgta cgaactagct
     3061 cccgagaaga cacagcctgc tctgctcagc acattcagtg gagaccgacg cttttctcga
     3121 tttggtagtg ttctgcacct gactgacctg gatgatgatg gcttagatga aatcatcatg
     3181 gctgccccac tgaggataac agatgtcact tccggactgc tagggggaga agatgggcga
     3241 gtgtacattt ataatggcat gtataccacc ttaggtgaca tgaccggcaa atgcaagtcc
     3301 tggatgactc catgtccgga ggagaaggca cagtatgtac taacttctcc agaggcaagt
     3361 tcaaggtttg ggagctcgct ggtgtccgtg cggtctaagg gaaggaacca agtggttgtt
     3421 gctgctggaa ggagttcttg gggagcccgg ctctctgggg cacttcatgt ctacagcttc
     3481 agctcagacc tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3541 tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc agcctcgact
     3601 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3661 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3721 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3781 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3841 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3901 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3961 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4021 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4081 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4141 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4201 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4261 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4321 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4381 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4441 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4501 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4561 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4621 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4681 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4741 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4801 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4861 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4921 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4981 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5041 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5101 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5161 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5221 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5281 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5341 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5401 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5461 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5521 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5581 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5641 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5761 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5821 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5881 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5941 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6001 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6061 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6661 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6781 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7981 aaaagtgcca cctgacgtc
//
