LOCUS       V020210                 8525 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020210
VERSION     V020210
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             941..4015
                     /label="NAT10(NM_024662)"
                     /note="NAT10(NM_024662)"
                     /gene="NAT10"
     CDS             4022..4087
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4125..4349
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4395..4823
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4837..5166
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5233..6027
                     /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    6201..6322
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6371..6387)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6395..6411
                     /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(6419..6449)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6464..6485
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6773..7358)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7529..8389)
                     /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(8390..8494)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcatcgga aaaaggtgga
      961 taaccgaatc cggattctca ttgagaatgg agtagctgag cggcaaagat ctctctttgt
     1021 tgtagttggg gatcgaggaa aagatcaggt ggtaatactt catcacatgt tatccaaagc
     1081 aactgtgaag gctcggcctt cagtgctgtg gtgttataag aaagagctgg ggtttagcag
     1141 tcaccggaag aaaagaatgc gacagctgca gaagaaaata aagaatggaa cactgaacat
     1201 aaagcaggac gacccctttg aactcttcat agcagccaca aacattcgct actgctacta
     1261 caacgagacc cacaagatcc tgggcaatac cttcggcatg tgtgtgctgc aggattttga
     1321 agccttaact ccaaacttgc tggccaggac tgtagaaaca gtggaaggtg gtgggctagt
     1381 ggtcatcctc ctacggacca tgaactcact caagcaattg tacacagtga ctatggatgt
     1441 gcattccagg tacagaactg aggcccatca ggatgtggtg ggaagattta atgaaaggtt
     1501 tattctgtct ctggcctctt gtaagaagtg tctcgtcatt gatgaccagc tcaacatcct
     1561 gcccatctcc tcccacgttg ccaccatgga ggccctgcct ccccagactc cggatgagag
     1621 tcttggtcct tctgatctgg agctgaggga gttgaaggag agcttgcagg acacccagcc
     1681 tgtgggtgtg ttggtggact gctgtaagac tctagaccag gccaaagctg tcttgaaatt
     1741 tatcgagggc atctctgaaa agaccctgag gagtactgtt gcactcacag ctgctcgagg
     1801 acggggaaaa tctgcagccc tgggattggc gattgctggg gcggtggcat ttgggtactc
     1861 caatatcttt gttacctccc caagccctga taacctccat actctgtttg aatttgtatt
     1921 taaaggattt gatgctctgc aatatcagga acatctggat tatgagatta tccagtctct
     1981 aaatcctgaa tttaacaaag cagtgatcag agtgaatgta tttcgagaac acaggcagac
     2041 tattcagtat atacatcctg cagatgctgt gaagctgggc caggctgaac tagttgtgat
     2101 tgatgaagct gccgccatcc ccctcccctt ggtgaagagc ctacttggcc cctaccttgt
     2161 tttcatggca tccaccatca atggctatga gggcactggc cggtcactgt ccctcaagct
     2221 aattcagcag ctccgtcaac agagcgccca gagccaggtc agcaccactg ctgagaataa
     2281 gaccacgacg acagccagat tggcatcagc gcggacactg tatgaggttt ccctccagga
     2341 gtcaatccga tacgcccctg gggatgcagt ggagaagtgg ctgaatgact tgctgtgcct
     2401 ggattgcctc aacatcactc ggatagtctc aggctgcccc ttgcctgaag cttgtgaact
     2461 gtactatgtt aatagagata ccctcttttg ctaccacaag gcctctgaag ttttcctcca
     2521 acggcttatg gccctctacg tggcttctca ctacaagaac tctcccaatg atctccagat
     2581 gctctccgat gcacctgctc accatctctt ctgccttctg cctcccgtgc cccccaccca
     2641 gaatgccctt ccagaagtgc ttgctgttat ccaggtgtgc cttgaagggg agatttctcg
     2701 ccagtccatc ttgaacagtc tgtctcgagg caagaaggct tcaggggacc tgattccatg
     2761 gacagtgtca gaacagttcc aagatccaga ctttggtggt ctgtctggtg gaagggtcgt
     2821 tcgcattgct gttcacccag attatcaagg gatgggctat ggcagccgtg ctctgcagct
     2881 gctgcagatg tactatgaag gcaggtttcc ttgtctggag gaaaaggtcc ttgagacacc
     2941 acaggaaatt cacaccgtaa gcagcgaggc tgtcagcttg ttggaagagg tcatcactcc
     3001 ccggaaggac ctgcctcctt tactcctcaa attgaatgag aggcctgccg aacgcctgga
     3061 ttacctgggt gtttcctatg gcttgacccc caggctcctc aagttctgga aacgagctgg
     3121 atttgttcct gtttatctga gacagacccc gaatgacctg accggagagc actcgtgcat
     3181 catgctgaag acgctcactg atgaggatga ggctgaccag ggaggctggc ttgcagcctt
     3241 ctggaaagat ttccgacggc ggttcctagc cttgctctcc taccagttca gtaccttctc
     3301 tccttccctg gctctgaaca tcattcagaa caggaacatg gggaagccag cccagcctgc
     3361 cctgagccgg gaggagctgg aagcactctt cctcccctat gacctgaagc ggctggagat
     3421 gtattcacgg aatatggtgg actatcacct catcatggac atgatcccgg ccatctctcg
     3481 catctatttc ctgaaccagc tgggggacct ggccctgtct gcggctcagt cggctcttct
     3541 cttggggatt ggcctgcagc ataagtctgt ggaccagctg gaaaaggaga ttgagctgcc
     3601 ctcgggccag ttgatgggac ttttcaaccg gatcatccgc aaagttgtga agctatttaa
     3661 tgaagttcag gaaaaggcca ttgaggagca gatggtggca gcgaaggatg tggtcatgga
     3721 gcccacgatg aagaccctca gtgacgacct agatgaagca gcaaaggaat ttcaggagaa
     3781 acacaagaag gaagtaggga agctgaagag catggacctc tctgaataca taatccgtgg
     3841 ggacgatgaa gagtggaatg aagttttgaa caaagctggg ccgaacgcct cgatcatcag
     3901 cctgaaaagt gacaagaaaa ggaagttaga ggccaaacaa gaacccaaac agagcaagaa
     3961 gttgaagaac agagagacaa agaacaaaaa agatatgaaa ctgaagcgga agaaactcga
     4021 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     4081 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     4141 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4201 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4261 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4321 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4381 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4441 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4501 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4561 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4621 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4681 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4741 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4801 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4921 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4981 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5041 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5101 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5161 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     5221 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5281 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5341 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5401 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5461 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5521 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5581 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5641 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5701 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5761 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5821 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5881 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5941 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6001 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6061 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6121 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6181 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6241 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6301 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6361 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6421 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6481 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6541 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6781 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6841 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6901 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6961 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7021 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7081 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7141 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7201 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7261 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     7321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7501 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7741 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7801 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8161 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8521 acgtc
//
