LOCUS       V032324                 8359 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032324
VERSION     V032324
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             874..1590
                     /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             1642..3849
                     /label="BACH1(NM_206866)"
                     /note="BACH1(NM_206866)"
                     /gene="BACH1"
     polyA_signal    3926..4150
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4196..4624
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4638..4967
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5034..5828
                     /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    6002..6123
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6172..6188)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6196..6212
                     /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(6220..6250)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6265..6286
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6574..7159)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7330..8190)
                     /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(8191..8295)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgtctctg agtgagaact cggtttttgc ctatgaatct
     1681 tctgtgcata gcaccaatgt tttactcagc cttaatgacc agcggaagaa agatgtgctg
     1741 tgcgatgtca ccatctttgt ggagggacag cggttccgcg ctcaccggtc cgtgctggcg
     1801 gcatgcagca gttacttcca ctcaagaatc gtaggccagg ctgatggaga gctgaacatt
     1861 actcttccag aagaggtgac agttaaagga tttgaacctt taattcagtt tgcctacact
     1921 gctaaactga ttttaagtaa agagaatgtg gatgaagtgt gcaaatgtgt ggagttttta
     1981 agtgtacata atattgagga atcctgcttt cagtttctga aatttaagtt tttggactcc
     2041 actgcagacc agcaagaatg cccaagaaaa aaatgctttt catcacactg tcagaaaaca
     2101 gaccttaaac tttcactttt ggaccagagg gatctagaaa ctgatgaagt ggaggaattt
     2161 ctggaaaata aaaatgttca gactcctcag tgtaaactcc gcaggtatca aggaaatgca
     2221 aaagcctcac ctcctctaca agacagtgcc agtcagacat atgagtccat gtgcttagag
     2281 aaggatgctg ctctggcctt gccttcttta tgccccaaat acagaaaatt ccaaaaagca
     2341 tttggaactg acagagtccg tactggggaa tctagtgtca aagacattca tgcttctgtt
     2401 cagccaaatg aaaggtctga aaatgaatgc ctgggaggag tcccggagtg tagagatttg
     2461 caggtgatgt taaaatgtga cgaaagtaaa ttagcaatgg aacctgaaga aacgaagaaa
     2521 gatcctgctt ctcagtgccc aactgaaaaa tcagaagtga ctcctttccc ccacaattct
     2581 tccatagacc ctcatggact ttattctttg tctcttttac acacatatga ccaatatggt
     2641 gacttgaatt ttgctggtat gcaaaacaca acagtgttaa cagaaaagcc tttgtcaggt
     2701 acagacgtcc aagaaaaaac atttggtgaa agtcaggatt tacctttgaa atccgacttg
     2761 ggcaccaggg aagatagtag tgttgcatct agtgatagga gtagtgtgga gcgagaagtg
     2821 gcagaacacc tagcaaaagg cttctggagt gacatttgca gcacggacac tccttgccaa
     2881 atgcagttat cacctgctgt ggccaaagat ggctcagaac agatctcaca gaaacggtct
     2941 gagtgtccgt ggttaggtat caggattagt gagagcccag aaccaggtca aaggactttc
     3001 acaacattaa gttctgtcaa ctgccctttt ataagtactc tgagtactga aggctgttca
     3061 agcaatttgg aaattggaaa cgatgattat gtttcagaac cccagcaaga accttgccca
     3121 tatgcttgtg tcattagctt gggagacgac tctgagacgg acaccgaagg agacagtgaa
     3181 tcctgttcag ccagagaaca agaatgtgag gtaaaactgc cattcaatgc acaacggata
     3241 atttcactgt ctcgaaatga ttttcagtcc ttgttgaaaa tgcacaagct tactccagaa
     3301 cagctggatt gtatccatga tattcgaaga agaagtaaaa acagaattgc tgcacagcgc
     3361 tgtcgcaaga gaaaacttga ctgtatacag aatcttgaat cagaaattga gaagctgcaa
     3421 agtgaaaagg agagcttgtt gaaggaaaga gatcacattt tgtcaactct gggtgagaca
     3481 aagcagaacc taactggact ttgccagaaa gtttgtaaag aagcagctct gagtcaagaa
     3541 caaatacaga tactcgccaa gtactcagct gcagattgcc cactttcatt tttaatttct
     3601 gaaaaagata aaagtactcc tgatggtgaa ctggcgttac catcaatttt cagtttatct
     3661 gaccggcctc cagcagtgct gcctccctgt gccagaggaa acagtgagcc tggctacgcg
     3721 cgagggcagg agtcccagca gatgtccaca gccacctctg agcaagctgg gcctgcggaa
     3781 cagtgtcgtc agagtggtgg gatctcagat ttctgtcagc agatgactga taaatgtact
     3841 actgatgagt aaggatcctg atatccagca cagtggcggc cgctcgagtc tagagggccc
     3901 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3961 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4021 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4081 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4141 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     4201 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4261 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4321 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4381 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4441 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4501 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4561 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4621 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4741 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4801 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4861 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4921 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4981 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5041 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5101 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5161 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5221 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5281 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5341 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5401 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5461 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5521 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5581 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5641 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5701 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5761 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5821 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5881 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5941 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6001 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6061 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6121 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6181 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6241 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6301 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6361 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6421 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6481 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6541 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6601 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6661 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6721 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6781 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6841 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6901 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6961 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7021 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7081 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     7141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7321 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7381 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7441 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7501 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7561 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7621 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7681 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7741 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7801 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7861 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7921 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7981 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8041 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8101 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8161 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8221 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8281 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8341 tctcccgatc ccctatggt
//
