LOCUS       V018249                 6556 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018249
VERSION     V018249
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..1302
                     /label="SUMO1(NM_003352)"
                     /note="SUMO1(NM_003352)"
                     /gene="SUMO1"
     polyA_signal    1315..1937
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        1979..2276
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2330..3124
                     /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    3776..3910
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(3916..3934)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3948..3964)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3972..3988
                     /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(3996..4026)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4041..4062
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4350..4938)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5109..5969)
                     /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(5970..6074)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6101..6529
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgtctgacca ggaggcaaaa
     1021 ccttcaactg aggacttggg ggataagaag gaaggtgaat atattaaact caaagtcatt
     1081 ggacaggata gcagtgagat tcacttcaaa gtgaaaatga caacacatct caagaaactc
     1141 aaagaatcat actgtcaaag acagggtgtt ccaatgaatt cactcaggtt tctctttgag
     1201 ggtcagagaa ttgctgataa tcatactcca aaagaactgg gaatggagga agaagatgtg
     1261 attgaagttt atcaggaaca aacggggggt cattcaacag tttaggcgga tcccgggtgg
     1321 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     1381 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     1441 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     1501 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     1561 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     1621 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     1681 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     1741 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta
     1801 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa
     1861 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag
     1921 tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg
     1981 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2041 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2101 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2161 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2221 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggagga
     2281 attgatcagc ttgggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2341 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2401 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2461 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     2521 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2581 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     2641 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     2701 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     2761 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     2821 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     2881 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     2941 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3001 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3061 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3121 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3181 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3241 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccggg
     3301 ctcgatcccc tcgcgagttg gttcagctgc tgcctgaggc tggacgacct cgcggagttc
     3361 taccggcagt gcaaatccgt cggcatccag gaaaccagca gcggctatcc gcgcatccat
     3421 gcccccgaac tgcaggagtg gggaggcacg atggccgctt tggtcgaccc ggacgggacg
     3481 ctcctgcgcc tgatacagaa cgaattgctt gcaggcatct catgagtgtg tcttcccgtt
     3541 ttccgcctga ggtcactgcg tggatggagc gctggcgcct gctgcgcgac ggcgagctgc
     3601 tcaccaccca ctcgccaagc tggaaccgta aaaaggccgc gttgctggcg tttttccata
     3661 ggctccgccg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3721 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3901 tgtctggatc aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt
     3961 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4021 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4081 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4141 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4201 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4261 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5041 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5101 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5161 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5221 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5281 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5341 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5401 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5461 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5521 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5581 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5641 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5701 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5761 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5821 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5881 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5941 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6001 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6061 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     6121 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6181 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6241 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6301 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6361 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6541 ttaacgctta caattt
//
