LOCUS       V039354                 8967 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039354
VERSION     V039354
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             955..3780
                     /label="USP33(NM_015017)"
                     /note="USP33(NM_015017)"
                     /gene="USP33"
     CDS             3817..4536
                     /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    4562..4786
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4832..5260
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5274..5603
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5670..6464
                     /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    6640..6761
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6810..6826)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6834..6850
                     /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(6858..6888)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6903..6924
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7212..7800)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7971..8831)
                     /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(8832..8936)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatgaca
      961 ggatcaaatt cacacataac gatattaacc ttaaaggtgt tacctcattt tgaaagtctt
     1021 gggaaacagg aaaaaattcc taacaaaatg tcagcttttc gaaatcattg tccacatttg
     1081 gattcagttg gtgaaataac aaaagaagat ttgatacaaa aatcccttgg tacttgtcag
     1141 gattgtaaag tccaaggacc aaatctttgg gcatgtctgg agaatagatg ttcatatgtt
     1201 ggctgtggtg aatcacaagt agatcacagc accatacatt ctcaggagac aaagcattat
     1261 ctaactgtga accttaccac tcttcgagta tggtgttatg cttgcagcaa agaagtattt
     1321 ttggatagga aattaggaac tcagccttca ttgcctcatg taagacaacc tcaccaaata
     1381 caagaaaaca gtgtccagga ttttaaaata cccagtaata caacattaaa aactcctctg
     1441 gttgccgtat ttgatgatct ggatatagaa gcggatgaag aagatgaact tagggccaga
     1501 ggtcttacag gtttgaaaaa tattggaaat acttgttaca tgaatgcagc tttgcaggct
     1561 ctttctaatt gcccaccttt gacacagttt tttcttgatt gtggaggact agctcgaaca
     1621 gataagaaac ctgccatttg taaaagttat ctcaaactaa tgacagagct gtggcataaa
     1681 agcaggccag gatctgttgt gcctactact ctgtttcaag gaattaaaac tgtaaatcca
     1741 acatttcggg ggtattctca gcaggatgct caagaattcc ttcgatgttt aatggatttg
     1801 cttcatgaag aattgaaaga gcaagtcatg gaagtagaag aagatccgca aaccataacc
     1861 actgaggaga caatggaaga agacaagagc cagtcggatg tagattttca gtcttgtgaa
     1921 tcttgtagca acagtgatag agcagaaaat gaaaatggct ctagatgctt ttctgaagat
     1981 aataatgaaa caacaatgtt aattcaggat gatgaaaaca attcagaaat gtcaaaggat
     2041 tggcaaaaag agaagatgtg caataagatt aataaagtaa attctgaagg cgaatttgat
     2101 aaagatagag actctatatc tgaaacagtc gacttaaaca accaggaaac tgtcaaagtg
     2161 caaatacaca gcagagcttc agaatatatc actgatgtcc attcgaatga cctgtctaca
     2221 ccacagatcc ttccatcaaa tgaaggtgtt aatccacgtt tatcggcaag ccctcctaaa
     2281 tcaggcaatt tgtggccagg attggcacca ccacacaaaa aagctcagtc tgcatctcca
     2341 aagagaaaaa aacagcacaa gaaatacaga agtgttattt cagacatatt tgatggaaca
     2401 atcattagtt cagtgcagtg tctgacttgt gacagggtgt ctgtaaccct cgagaccttt
     2461 caagatctgt ccttgccaat tcctggcaag gaagaccttg ctaagctgca ttcatcaagt
     2521 catccaactt ctatagtcaa agcaggatca tgtggcgaag catatgctcc acaagggtgg
     2581 atagcttttt tcatggaata tgtgaagagg tttgttgtct catgtgtccc tagctggttt
     2641 tggggtccag tagtaacctt gcaagattgt cttgctgcct tctttgccag agatgaacta
     2701 aaaggtgaca atatgtacag ttgtgaaaaa tgcaaaaagt tgagaaatgg agtgaagttt
     2761 tgtaaagtac aaaactttcc tgagattttg tgcatccacc ttaaaagatt cagacatgaa
     2821 ctaatgtttt ccaccaaaat cagtacccat gtttcatttc cgctagaagg cttggatctt
     2881 cagccatttc ttgctaagga tagtccagct caaattgtga catatgatct tctgtcagtc
     2941 atttgccatc atggaactgc aagtagtgga cactatatag cctactgccg aaacaatcta
     3001 aataatctct ggtatgaatt tgatgatcag agtgtcactg aagtttcaga atctactgta
     3061 caaaatgcag aagcttacgt tcttttctat aggaagagca gcgaagaggc acaaaaagag
     3121 aggagaagga tatcaaattt attgaacata atggaaccaa gcctccttca gttttatatt
     3181 tctcgacagt ggcttaataa atttaagacc tttgccgaac ctggccctat ttcaaataat
     3241 gactttcttt gtattcatgg aggtgttcct ccaagaaaag ctggttatat tgaagacctg
     3301 gttttgatgc tgcctcagaa catttgggat aacctatata gcaggtatgg tggaggacca
     3361 gctgtcaacc atctgtacat ttgtcatact tgccaaattg aggcggagaa aattgaaaaa
     3421 agaagaaaaa ctgaattgga aatttttatt cggcttaaca gagcgttcca aaaagaggac
     3481 tctccagcta ctttttattg catcagtatg cagtggttta gagaatggga aagttttgtg
     3541 aagggtaaag atggagatcc tccaggtcct attgacaata ctaagattgc agtcactaaa
     3601 tgtggtaatg tgatgcttag gcaaggagca gattctggcc agatttctga agaaacatgg
     3661 aattttctgc agtctattta tggtggaggg cctgaagtta tcctgcgacc tccggttgtt
     3721 catgttgatc cagatatact tcaagcagaa gaaaaaattg aagtagaaac tcggtctttg
     3781 ctcgagtcta gagggccctt cgggggtgga ggctctatgg tgagcaaggg cgaggagctg
     3841 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     3901 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3961 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     4021 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     4081 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     4141 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     4201 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     4261 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     4321 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     4381 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     4441 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4501 gggatcactc tcggcatgga cgagctgtac aagtaagttt aaacccgctg atcagcctcg
     4561 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4621 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4681 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4741 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4801 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5041 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5101 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5161 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5221 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5281 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5341 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5401 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5461 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5521 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5581 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5641 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5701 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5761 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5821 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5881 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5941 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6001 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6061 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6121 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6181 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6241 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6301 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6361 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6421 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6481 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6541 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6601 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6661 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6721 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6781 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6841 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6901 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7021 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7081 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7141 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7621 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7681 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7741 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7801 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7861 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7921 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7981 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8041 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8101 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8161 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8221 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8281 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8341 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8401 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8461 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8521 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8581 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8641 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8701 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8761 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8821 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8881 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8941 tttccccgaa aagtgccacc tgacgtc
//
