LOCUS       V033148                 6077 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033148
VERSION     V033148
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             941..1567
                     /label="TIMM23(NM_006327)"
                     /note="TIMM23(NM_006327)"
                     /gene="TIMM23"
     CDS             1574..1639
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1677..1901
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      1947..2375
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2389..2718
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2785..3579
                     /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    3753..3874
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3923..3939)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3947..3963
                     /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(3971..4001)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4016..4037
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4325..4910)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5081..5941)
                     /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(5942..6046)
                     /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 atccgccacc atggaaggag gcgggggaag
      961 cggcaacaaa accacagggg gattggccgg ctttttcgga gccggcggag caggttactc
     1021 gcacgcggat ttggctggcg tcccgctaac tggtatgaac cctctgtctc cttatttaaa
     1081 tgtggatcca cgatacctcg tgcaggatac agatgagttt attttaccta ccggagctaa
     1141 taaaacccgg ggcagatttg agctggcctt ctttacgatt ggaggatgtt gcatgacagg
     1201 ggctgcgttt ggtgcaatga atggtcttcg gctaggattg aaggaaaccc agaacatggc
     1261 ctggtccaaa ccaagaaatg tacagatttt gaatatggtg actaggcaag gggcactttg
     1321 ggctaatact ctaggttctc tggctttgct ctatagtgca tttggtgtca tcattgagaa
     1381 aacacgaggt gcagaagatg accttaacac agtagcagct ggaaccatga caggcatgtt
     1441 gtataaatgt acaggtggtc ttcgagggat agcacgaggt ggtctgacag gactaacact
     1501 taccagcctc tatgcactat ataataactg ggagcacatg aaaggctcct tgctccaaca
     1561 gtcactcctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     1621 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt
     1681 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1741 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1801 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1861 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1921 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1981 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2041 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2101 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2161 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2221 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2281 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2341 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2401 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2461 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2521 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2581 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2641 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2701 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2761 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2821 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2881 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2941 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3001 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3061 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3121 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3181 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3241 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3301 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3361 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3421 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3481 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3541 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3601 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3661 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3721 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3781 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3841 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3901 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4741 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6061 aagtgccacc tgacgtc
//
