LOCUS       V026043                 8769 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026043
VERSION     V026043
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             1660..4233
                     /label="USP5(NM_001098536)"
                     /note="USP5(NM_001098536)"
                     /gene="USP5"
     polyA_signal    4336..4560
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4606..5034
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5048..5377
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5444..6238
                     /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    6412..6533
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6582..6598)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6606..6622
                     /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(6630..6660)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6675..6696
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6984..7569)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7740..8600)
                     /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(8601..8705)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tggcggagct gagtgaggag
     1681 gcgctgctgt cagtattacc gacgatccgg gtccctaagg ctggagaccg ggtccacaaa
     1741 gacgagtgcg ccttctcctt cgacacgccg gagtctgagg ggggcctcta catctgtatg
     1801 aacacgtttc tgggctttgg gaaacagtat gtggagagac atttcaataa gaccggccag
     1861 cgagtctact tgcacctccg gcggacccgg cgcccgaaag aggaggaccc tgctacaggc
     1921 actggagacc caccccggaa gaagcccacg cggctggcta ttggtgttga aggcggattt
     1981 gaccttagcg aggagaagtt tgaattagac gaggatgtga agattgtcat tttgccagat
     2041 tacctggaga ttgcccggga tggactgggg ggactgcctg acattgtcag agatcgggtg
     2101 accagtgcag tggaggccct actgtcggcc gactcagcct cccgcaagca ggaggtgcag
     2161 gcatgggatg gggaagtacg gcaggtgtct aagcatgcct tcagcctcaa gcagttggac
     2221 aaccctgctc gaatccctcc ctgtggctgg aagtgctcca agtgtgacat gagagagaac
     2281 ctgtggctca acctgactga tggctccatc ctctgtgggc gacgctactt cgatggcagt
     2341 gggggcaaca accacgctgt ggagcactac cgagagacag gctacccgtt agctgtcaag
     2401 ctgggcacca tcacccctga tggagctgac gtgtactcat atgatgagga tgacatggtc
     2461 ctggacccca gcctggctga gcacctgtcc cacttcggca tcgacatgct gaagatgcag
     2521 aagacagaca agacgatgac tgagttggag atagacatga accagcggat tggtgaatgg
     2581 gagctgatcc aggagtcagg tgtgccactc aagcccctgt ttgggcctgg ctacacaggc
     2641 atccggaacc tgggtaacag ctgctacctc aactctgtgg tccaggtgct cttcagcatc
     2701 cctgacttcc agaggaagta tgtggataag ctggagaaga tcttccagaa tgccccgacg
     2761 gaccctaccc aggatttcag cacccaggtg gccaagctgg gccatggcct tctctccggg
     2821 gagtattcca agccagtacc ggagtcgggc gatggggagc gggtgccaga acagaaggaa
     2881 gttcaagatg gcattgcccc tcggatgttc aaggccctca tcggcaaggg ccaccctgaa
     2941 ttctccacca accggcagca ggatgcccag gagttcttcc ttcaccttat caacatggtg
     3001 gagaggaatt gccggagctc tgaaaatcct aatgaagtgt tccgcttctt ggtggaggaa
     3061 aagatcaagt gcctggccac agagaaggtg aagtacaccc agcgagttga ctacatcatg
     3121 cagctgcctg tgcccatgga tgcagccctt aacaaagagg agcttctgga gtacgaggag
     3181 aagaagcggc aagccgaaga ggagaagatg gcactgccag aactggttcg ggcccaggtg
     3241 cccttcagct cttgcctgga ggcctacggg gcccctgagc aggtcgatga cttctggagc
     3301 acggccctgc aggccaagtc agtagctgtc aagaccacac gatttgcctc attccctgac
     3361 tacctggtca tccagatcaa gaagttcacc ttcggcttag actgggtgcc caagaaactg
     3421 gatgtgtcca tcgagatgcc agaggagctc gacatctccc agttgagggg cacagggctg
     3481 cagcccggag aggaggagct gccagacatt gccccacccc tggtcactcc ggatgagccc
     3541 aaaggtagcc ttggtttcta tggcaacgaa gacgaagact ccttctgctc ccctcacttc
     3601 tcctctccga catcgcccat gctggatgaa tcagtcatca tccagctggt ggagatggga
     3661 ttccctatgg acgcctgccg caaagctgtc tactacacgg gcaacagcgg ggctgaggcc
     3721 gccatgaact gggtcatgtc acacatggat gatccagatt ttgcaaaccc cctcatcctg
     3781 cctggctcta gtgggccggg ctccacaagc gcagcagccg acccccctcc tgaggactgt
     3841 gtgaccacca ttgtctccat gggcttctcc cgggaccagg ccttgaaagc gctgcgggcc
     3901 acgaacaata gtttagaacg ggctgtggac tggatcttca gtcacattga cgacctggat
     3961 gctgaagctg ccatggacat ctcagagggc cgctcagctg ccgactccat ctctgagtct
     4021 gtgccagtgg gacctaaagt ccgggatggt cctggaaagt atcagctctt tgccttcatt
     4081 agtcacatgg gcacctctac catgtgtggt cactacgtct gccacatcaa gaaagaaggc
     4141 agatgggtga tctacaatga ccagaaagtg tgtgcctccg agaagccgcc caaggacctg
     4201 ggctacatct acttctacca gagagtggcc agctaaggat ccactagtcc agtgtggtgg
     4261 aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc
     4321 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4381 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4441 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4501 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4561 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4621 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4681 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4741 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4801 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4861 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4921 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4981 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     5041 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5101 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5221 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5281 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     5341 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     5401 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5461 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5521 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5581 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5641 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5701 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5761 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5821 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5881 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5941 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6001 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6061 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6121 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6181 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6241 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6301 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6361 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6421 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6481 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6541 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6601 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6661 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6721 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6781 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6841 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6901 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6961 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7381 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7501 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7681 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7861 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8161 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8341 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8701 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8761 ccctatggt
//
