LOCUS       V046991                 7254 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046991
VERSION     V046991
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..1934
                     /label="Idh3g(NM_008323)"
                     /note="Idh3g(NM_008323)"
                     /gene="Idh3g"
     CDS             1941..2006
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2030..2618
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2684..2732
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2934..3362
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3376..3705
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3772..4566
                     /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    4742..4863
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4912..4928)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4936..4952
                     /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(4960..4990)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5005..5026
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5314..5902)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6073..6933)
                     /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(6934..7038)
                     /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 gctgaaggtg gcgatagctg
      781 ctggcggtgc tgcaaaggca atgctcaagc caactctcct ctgccgtcct tgggaggttc
      841 tggctgccca tgtggccccc cgaaggagca tttcctcaca acaaacaatt cctccatctg
      901 ctaagtatgg tgggcggcat acagtgacta tgatcccagg ggatggcatc ggcccagagc
      961 tcatgttgca tgttaagtct gtattcaggc atgcatgtgt gccggtggac tttgaagagg
     1021 tgcatgtaag ctccaacgct gatgaggagg acatccgcaa tgccatcatg gccatccgcc
     1081 ggaaccgtgt ggccctgaag ggcaacattg aaacaaatca taacctgcca ccatcccaca
     1141 aatctcgaaa caacatcctt cgcaccagcc tagacctcta tgccaacgtc atccactgta
     1201 agagcctgcc aggagtggtg acccggcaca aggacataga catcctcatt gtacgggaaa
     1261 acacagaagg cgagtacagc agcctggagc atgagagcgt agcaggagtg gtggagagct
     1321 tgaagattat caccaaagcc aagtccctgc gcattgctga atatgctttc aagctggccc
     1381 aggagagtgg gcgtaagaaa gtgacggctg tgcacaaggc caacatcatg aaactgggtg
     1441 atggactctt cctccagtgc tgcagggaag tagcagccca ctaccctcag atcacctttg
     1501 acagcatgat tgtagacaac acaacaatgc agctggtatc ccggcctcag cagtttgatg
     1561 tcatggtgat gcctaatctc tatggtaaca ttgtcaacaa cgtctgtgca gggctagttg
     1621 gaggcccagg ccttgtggct ggggccaact atggccatgt gtatgcagta ttcgagacag
     1681 ctacaaggaa cacaggcaaa agtattgcca ataagaacat tgctaacccg actgccacac
     1741 tgctagcaag ctgcatgatg ctagaccacc tcaagctcca ctcctatgcc acttccatcc
     1801 gcaaagctgt cttagcatcc atggacaatg aaaatatgca taccccagat attggaggcc
     1861 agggcaccac atcccaagcc atccaggaca tcattcgtca tatccgcatc attaatggac
     1921 gggctgtgga ggctggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     1981 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     2041 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2101 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2161 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2221 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2281 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2341 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2401 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     2461 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2521 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2581 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     2641 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     2701 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     2761 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     2821 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     2881 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     2941 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3001 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3061 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     3121 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3181 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3241 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3301 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3361 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3421 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3481 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3541 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3601 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3661 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3721 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3781 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3841 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3901 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3961 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4021 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4081 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4141 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4201 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4261 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4321 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4381 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4441 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4501 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4561 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     4621 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4681 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4741 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4861 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4921 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4981 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5041 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5101 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     5521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6121 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6181 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6241 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6301 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6361 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6421 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6481 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6541 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6601 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6661 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6721 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6781 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6841 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6901 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6961 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7021 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7081 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7141 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7201 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
