LOCUS       V030824                 8179 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030824
VERSION     V030824
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2901
                     /label="THOC1(NM_005131)"
                     /note="THOC1(NM_005131)"
                     /gene="THOC1"
     CDS             2917..3633
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3634..3657
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3676..3699
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     polyA_signal    3746..3970
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4016..4444
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4458..4787
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4854..5648
                     /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    5822..5943
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5992..6008)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6016..6032
                     /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(6040..6070)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6085..6106
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6394..6979)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7150..8010)
                     /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(8011..8115)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtctccga cgccgccgct cttcagtttg
      961 cccgaagcgc ggacgcggtt tacgaagtct accagagagg ccttgaacaa caaaaacatc
     1021 aagccattgt taagtacctt cagccaggta cctggcagtg aaaatgaaaa aaaatgtacc
     1081 cttgaccaag ctttcagagg tattctagaa gaagaaatta taaatcattc atcatgtgaa
     1141 aacgttttag ctattatttc tcttgctatt gggggagtaa ctgaaggtat ttgtaccgca
     1201 tctacacctt ttgtattgtt gggagatgtt ttggattgtc ttcctttgga tcagtgtgac
     1261 acaatattca cttttgtgga aaaaaatgtt gctacttgga aatcaaatac attctattct
     1321 gctgggaaaa attacttact acgtatgtgc aatgatctcc taagaagatt gtctaaatcc
     1381 cagaatacag tcttctgtgg acggattcag ctctttttgg ccaggctttt ccctctgtct
     1441 gagaaatcag gtcttaactt gcagagtcag tttaatctgg aaaatgtcac tgttttcaat
     1501 acaaatgagc aggaaagcac cctgggtcag aagcacactg aagatagaga agaaggaatg
     1561 gatgtagaag aaggcgaaat gggagacgag gaagctccaa caacgtgctc tattccaatt
     1621 gattacaacc tgtatcgaaa attctggtca cttcaggatt acttcaggaa ccctgtgcaa
     1681 tgctatgaga agatttcatg gaaaactttt ctcaagtatt ctgaagaagt tttagctgtt
     1741 tttaagagtt ataaattaga tgatactcag gcctcaagaa aaaagatgga agaattgaaa
     1801 acaggaggag aacatgtata ttttgcaaaa tttttaacaa gtgaaaagct gatggattta
     1861 caactgagtg acagtaactt tcgtcgacac atcctgttgc agtatctcat tttattccaa
     1921 tatctcaagg ggcaggtcaa attcaaaagt tcaaactatg ttttaactga tgagcaatca
     1981 ctttggattg aagatactac aaaatcagtt tatcaactac tatctgaaaa cccccccgat
     2041 ggagaaagat tttcaaagat ggtagagcat atattaaaca ctgaagaaaa ctggaactcg
     2101 tggaaaaatg aaggttgccc aagttttgtg aaagaaagaa catcagatac caaacctacg
     2161 agaataattc ggaagagaac agcacccgag gacttcctag ggaaaggacc caccaaaaaa
     2221 attctgatgg gaaatgagga gttaacaagg ctttggaatc tttgccctga taatatggaa
     2281 gcctgtaaat cagagacaag ggaacacatg cccactttgg aggaattctt tgaagaagcc
     2341 attgaacagg cagaccctga aaatatggtg gaaaatgaat ataaggctgt gaacaattca
     2401 aattatggtt ggagagccct gagactatta gcacggagaa gccctcactt cttccagcca
     2461 accaaccagc agtttaaaag tttaccagaa tatcttgaaa atatggtaat aaagctagcc
     2521 aaggaattac cgcctccttc tgaagaaata aaaacaggtg aggatgaaga tgaggaagat
     2581 aatgatgctc tactgaagga aaatgaaagt cctgatgttc ggcgagacaa acctgtaaca
     2641 ggagaacaaa tagaggtatt tgccaacaag ctgggtgaac aatggaagat tctggctccc
     2701 tacttggaaa tgaaagactc agaaattagg cagattgagt gtgacagtga agacatgaag
     2761 atgagagcta agcagctcct ggttgcctgg caagatcaag agggagttca tgcaacacct
     2821 gagaatctga ttaatgcact gaataagtct ggattaagtg accttgcaga aagtctaact
     2881 aatgacaatg agacaaatag tggaggaggg ggaagcatgg tgagcaaggg cgaggagctg
     2941 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     3001 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3061 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     3121 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3181 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3241 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3301 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3361 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3421 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3481 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3541 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3601 gggatcactc tcggcatgga cgagctgtac aagtggagcc acccgcagtt cgagaagggc
     3661 ggcggcagca gcgcctggag ccacccgcag ttcgagaagt gactcgagtc tagagggccc
     3721 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3781 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3841 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3901 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3961 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     4021 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4081 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4141 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4201 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4261 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4321 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4381 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4441 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4501 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4561 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4621 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4681 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4741 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4801 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4861 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4921 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4981 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5041 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5101 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5161 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5221 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5281 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5341 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5401 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5461 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5521 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5581 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5641 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5701 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5761 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5821 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5881 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5941 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6001 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6061 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6121 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6181 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6301 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6901 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6961 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7021 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7081 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7141 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7201 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7261 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7321 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7381 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7441 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7501 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7561 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7621 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7681 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7741 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7801 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7861 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7921 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7981 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8041 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8101 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8161 tctcccgatc ccctatggt
//
