LOCUS       V028773                 9249 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028773
VERSION     V028773
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..4716
                     /label="NAT10(NM_024662)"
                     /note="NAT10(NM_024662)"
                     /gene="NAT10"
     CDS             4723..4740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    4816..5040
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5086..5514
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5528..5857
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5924..6718
                     /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    6892..7013
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7062..7078)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7086..7102
                     /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(7110..7140)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7155..7176
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7464..8049)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8220..9080)
                     /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(9081..9185)
                     /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 catgcatcgg aaaaaggtgg ataaccgaat ccggattctc
     1681 attgagaatg gagtagctga gcggcaaaga tctctctttg ttgtagttgg ggatcgagga
     1741 aaagatcagg tggtaatact tcatcacatg ttatccaaag caactgtgaa ggctcggcct
     1801 tcagtgctgt ggtgttataa gaaagagctg gggtttagca gtcaccggaa gaaaagaatg
     1861 cgacagctgc agaagaaaat aaagaatgga acactgaaca taaagcagga cgaccccttt
     1921 gaactcttca tagcagccac aaacattcgc tactgctact acaacgagac ccacaagatc
     1981 ctgggcaata ccttcggcat gtgtgtgctg caggattttg aagccttaac tccaaacttg
     2041 ctggccagga ctgtagaaac agtggaaggt ggtgggctag tggtcatcct cctacggacc
     2101 atgaactcac tcaagcaatt gtacacagtg actatggatg tgcattccag gtacagaact
     2161 gaggcccatc aggatgtggt gggaagattt aatgaaaggt ttattctgtc tctggcctct
     2221 tgtaagaagt gtctcgtcat tgatgaccag ctcaacatcc tgcccatctc ctcccacgtt
     2281 gccaccatgg aggccctgcc tccccagact ccggatgaga gtcttggtcc ttctgatctg
     2341 gagctgaggg agttgaagga gagcttgcag gacacccagc ctgtgggtgt gttggtggac
     2401 tgctgtaaga ctctagacca ggccaaagct gtcttgaaat ttatcgaggg catctctgaa
     2461 aagaccctga ggagtactgt tgcactcaca gctgctcgag gacggggaaa atctgcagcc
     2521 ctgggattgg cgattgctgg ggcggtggca tttgggtact ccaatatctt tgttacctcc
     2581 ccaagccctg ataacctcca tactctgttt gaatttgtat ttaaaggatt tgatgctctg
     2641 caatatcagg aacatctgga ttatgagatt atccagtctc taaatcctga atttaacaaa
     2701 gcagtgatca gagtgaatgt atttcgagaa cacaggcaga ctattcagta tatacatcct
     2761 gcagatgctg tgaagctggg ccaggctgaa ctagttgtga ttgatgaagc tgccgccatc
     2821 cccctcccct tggtgaagag cctacttggc ccctaccttg ttttcatggc atccaccatc
     2881 aatggctatg agggcactgg ccggtcactg tccctcaagc taattcagca gctccgtcaa
     2941 cagagcgccc agagccaggt cagcaccact gctgagaata agaccacgac gacagccaga
     3001 ttggcatcag cgcggacact gtatgaggtt tccctccagg agtcaatccg atacgcccct
     3061 ggggatgcag tggagaagtg gctgaatgac ttgctgtgcc tggattgcct caacatcact
     3121 cggatagtct caggctgccc cttgcctgaa gcttgtgaac tgtactatgt taatagagat
     3181 accctctttt gctaccacaa ggcctctgaa gttttcctcc aacggcttat ggccctctac
     3241 gtggcttctc actacaagaa ctctcccaat gatctccaga tgctctccga tgcacctgct
     3301 caccatctct tctgccttct gcctcccgtg ccccccaccc agaatgccct tccagaagtg
     3361 cttgctgtta tccaggtgtg ccttgaaggg gagatttctc gccagtccat cttgaacagt
     3421 ctgtctcgag gcaagaaggc ttcaggggac ctgattccat ggacagtgtc agaacagttc
     3481 caagatccag actttggtgg tctgtctggt ggaagggtcg ttcgcattgc tgttcaccca
     3541 gattatcaag ggatgggcta tggcagccgt gctctgcagc tgctgcagat gtactatgaa
     3601 ggcaggtttc cttgtctgga ggaaaaggtc cttgagacac cacaggaaat tcacaccgta
     3661 agcagcgagg ctgtcagctt gttggaagag gtcatcactc cccggaagga cctgcctcct
     3721 ttactcctca aattgaatga gaggcctgcc gaacgcctgg attacctggg tgtttcctat
     3781 ggcttgaccc ccaggctcct caagttctgg aaacgagctg gatttgttcc tgtttatctg
     3841 agacagaccc cgaatgacct gaccggagag cactcgtgca tcatgctgaa gacgctcact
     3901 gatgaggatg aggctgacca gggaggctgg cttgcagcct tctggaaaga tttccgacgg
     3961 cggttcctag ccttgctctc ctaccagttc agtaccttct ctccttccct ggctctgaac
     4021 atcattcaga acaggaacat ggggaagcca gcccagcctg ccctgagccg ggaggagctg
     4081 gaagcactct tcctccccta tgacctgaag cggctggaga tgtattcacg gaatatggtg
     4141 gactatcacc tcatcatgga catgatcccg gccatctctc gcatctattt cctgaaccag
     4201 ctgggggacc tggccctgtc tgcggctcag tcggctcttc tcttggggat tggcctgcag
     4261 cataagtctg tggaccagct ggaaaaggag attgagctgc cctcgggcca gttgatggga
     4321 cttttcaacc ggatcatccg caaagttgtg aagctattta atgaagttca ggaaaaggcc
     4381 attgaggagc agatggtggc agcgaaggat gtggtcatgg agcccacgat gaagaccctc
     4441 agtgacgacc tagatgaagc agcaaaggaa tttcaggaga aacacaagaa ggaagtaggg
     4501 aagctgaaga gcatggacct ctctgaatac ataatccgtg gggacgatga agagtggaat
     4561 gaagttttga acaaagctgg gccgaacgcc tcgatcatca gcctgaaaag tgacaagaaa
     4621 aggaagttag aggccaaaca agaacccaaa cagagcaaga agttgaagaa cagagagaca
     4681 aagaacaaaa aagatatgaa actgaagcgg aagaaaaagc ttcatcacca tcatcaccat
     4741 taggatcctg atatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc
     4801 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4861 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4921 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4981 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5041 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     5101 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5161 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5221 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5281 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5341 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5401 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5461 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     5521 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5581 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5641 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5701 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5761 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     5821 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     5881 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5941 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6001 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6061 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     6121 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6181 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6241 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6301 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6361 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6421 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6481 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6541 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6601 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6661 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6721 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6781 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6841 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6901 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6961 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7021 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7081 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7141 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7201 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7261 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7321 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7381 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7441 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7501 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7561 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7621 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7681 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7741 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7801 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7861 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7921 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7981 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8041 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8101 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8161 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8221 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8281 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8341 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8401 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8461 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8521 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8581 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8641 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8701 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8761 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8821 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8881 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8941 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9001 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9061 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9121 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9181 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     9241 ccctatggt
//
