LOCUS       V030406                 8862 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030406
VERSION     V030406
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             927..3665
                     /label="CUL4B(NM_003588)"
                     /note="CUL4B(NM_003588)"
                     /gene="CUL4B"
     CDS             3705..4424
                     /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"
     polyA_signal    4462..4686
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4732..5160
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5174..5503
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5570..6364
                     /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    6538..6659
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6708..6724)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6732..6748
                     /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(6756..6786)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6801..6822
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7110..7695)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7866..8726)
                     /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(8727..8831)
                     /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 gtttaaagcc actgggatcc gccaccatga tgtcacagtc atctggatca ggagatggga
      961 atgatgatga ggctactacc tctaaagacg gtggtttttc ttcccccagt ccctcagctg
     1021 ctgctgctgc tcaggaggtc agatctgcca ctgatggtaa taccagcacc actccgccca
     1081 cctctgccaa gaagagaaag ttaaacagca gcagcagtag cagcagtaac agtagtaacg
     1141 agagagaaga ctttgattcc acctcttcct cctcttccac tcctccttta caacccaggg
     1201 attcggcatc cccttcaacc tcgtccttct gcctgggggt ttcagtggct gcttccagcc
     1261 acgtaccgat acagaagaag ctgcgttttg aagacaccct ggagtttgta gggtttgatg
     1321 cgaagatggc tgaggaatcc tcctcctcct cctcctcatc ttcaccaact gctgcaacat
     1381 ctcagcagca gcaacttaaa aataagagta tattaatctc ttctgtggct tcggtgcatc
     1441 atgcaaacgg cctagccaaa tcttctacca ccgtctctag ctttgctaac agcaaacctg
     1501 gctctgctaa gaagttagtg atcaagaact ttaaagataa gcctaaatta ccagaaaact
     1561 acacagatga aacctggcaa aaactgaaag aagcagtgga agctattcag aatagtactt
     1621 caattaagta caatttagaa gaactctacc aggctgtaga aaatctctgt tcttacaaga
     1681 tttctgcaaa cttgtacaaa cagctgagac agatctgcga agatcacatc aaagcacaga
     1741 ttcatcaatt cagagaggat tcattggata gcgttctttt tttaaagaag attgatagat
     1801 gctggcaaaa ccattgcaga caaatgatca tgatcaggag catttttttg tttctggata
     1861 gaacttacgt tcttcagaat tcaatgctac cctccatttg ggacatggga ctggagttat
     1921 ttagggctca tattataagt gatcagaaag tgcagaataa gacaattgat ggcattcttc
     1981 tcttgattga gagggaaagg aatggtgaag caattgatag aagtttactt cgaagccttt
     2041 taagcatgct gtctgatttg caaatttatc aagattcttt tgaacaacga tttttggaag
     2101 aaactaaccg gctctatgca gctgaaggcc aaaaattaat gcaagaaaga gaggttcctg
     2161 aatatctaca tcatgttaac aaacgtctag aagaagaagc agacagactt attacttact
     2221 tagatcagac cacccagaag tcattaattg ctactgtaga aaaacaactt ctaggtgaac
     2281 acttaacagc aattcttcag aaaggtttaa ataacctcct tgatgaaaac cgaattcaag
     2341 atttgtctct tctgtatcag ctcttcagta gagttcgagg tggagttcag gttcttttgc
     2401 agcagtggat cgaatatatc aaggcatttg gcagcactat tgtaattaat cctgaaaaag
     2461 ataaaaccat ggttcaagaa ttgctggatt ttaaagataa ggttgaccat ataattgata
     2521 tctgctttct gaagaatgag aaatttatca atgccatgaa agaagcattt gaaacgttca
     2581 ttaacaaaag accaaataaa ccagctgaac ttatagctaa gtatgtagat tcaaaacttc
     2641 gtgcaggcaa caaagaagct acagatgaag aacttgagaa aatgttggat aaaattatga
     2701 tcatatttag atttatctat ggcaaggatg tttttgaggc cttctataag aaagatttag
     2761 ccaagcgcct gttagtcgga aagagtgcat ctgtagatgc tgaaaaatca atgctgtcca
     2821 aacttaaaca tgaatgcgga gctgctttca ccagcaaact tgaaggaatg tttaaagaca
     2881 tggaactttc taaagacatc atgattcagt tcaaacagta tatgcagaat cagaatgttc
     2941 cgggaaatat tgagttaact gtgaatatcc tgacaatggg ctattggccg acatatgtgc
     3001 ctatggaagt tcatttacca ccagagatgg taaaacttca ggagattttc aagacatttt
     3061 acctaggcaa acatagtggc aggaaacttc agtggcagtc aaccctagga cactgtgtgt
     3121 taaaagcaga atttaaagag ggtaaaaagg aactccaggt ctctcttttt caaacactgg
     3181 tgctgctaat gtttaatgag ggagaggagt tcagtttaga agagatcaag caggcaactg
     3241 gaatagagga tggagagtta aggagaacac tgcagtcatt agcctgtggc aaagctagag
     3301 ttctggcgaa aaatccaaag ggcaaagaca ttgaagatgg tgacaagttc atttgtaatg
     3361 atgatttcaa acataaactt ttcaggataa agatcaatca aatccagatg aaagaaacgg
     3421 ttgaagaaca agcaagcact acagaaagag tatttcaaga cagacagtat caaattgatg
     3481 ctgcaattgt tcgaattatg aagatgagaa agacacttag ccacaatctc cttgtttcag
     3541 aagtgtacaa ccagttgaaa tttccagtaa agcctgctga tcttaagaag agaatagaat
     3601 ctttaattga ccgggactac atggaaagag ataaagaaaa tccaaaccag tacaactata
     3661 ttgcaggctc gagtggcggt ggaggaagcg gcggaggtgg atctatggtg agcaagggcg
     3721 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3781 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3841 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     3901 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3961 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4021 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4081 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4141 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     4201 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4261 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4321 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4381 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtagtctaga gggcccgttt
     4441 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4501 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4561 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4621 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4681 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     4741 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4801 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4861 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4921 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4981 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5041 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5101 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5161 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5221 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5281 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5341 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5401 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     5461 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     5521 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5581 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5641 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5701 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5761 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5821 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5881 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5941 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6001 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6061 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     6121 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6181 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6241 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6301 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6361 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6421 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6481 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     6541 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6601 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6661 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6721 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6781 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6841 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6901 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6961 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7021 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7081 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7141 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7201 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7261 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7321 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7381 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7441 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7501 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7561 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7621 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga
     7681 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7741 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7801 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8461 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8821 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
