LOCUS       V031447                 6559 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031447
VERSION     V031447
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..1308
                     /label="SUMO3(NM_006936)"
                     /note="SUMO3(NM_006936)"
                     /gene="SUMO3"
     polyA_signal    1318..1940
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        1982..2279
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2333..3127
                     /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    3779..3913
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(3919..3937)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3951..3967)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3975..3991
                     /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(3999..4029)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4044..4065
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4353..4941)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5112..5972)
                     /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(5973..6077)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6104..6532
                     /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 tgtccgagga gaagcccaag
     1021 gagggtgtga agacagagaa tgaccacatc aacctgaagg tggccgggca ggacggctcc
     1081 gtggtgcagt tcaagatcaa gaggcacacg ccgctgagca agctgatgaa ggcctactgc
     1141 gagaggcagg gcttgtcaat gaggcagatc agattcaggt tcgacgggca gccaatcaat
     1201 gaaactgaca ctccagcaca gctggagatg gaggacgagg acaccatcga cgtgttccag
     1261 cagcagacgg gaggtgtgcc ggagagcagc ctggcagggc acagtttcta ggatcccggg
     1321 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     1381 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     1441 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     1501 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     1561 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     1621 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     1681 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     1741 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     1801 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     1861 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     1921 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta
     1981 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2041 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2101 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2161 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2221 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2281 ggaattgatc agcttgggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     2341 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     2401 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2461 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     2521 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     2581 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2641 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2701 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2761 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2821 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     2881 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2941 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3001 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3061 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3121 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     3181 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3241 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     3301 gggctcgatc ccctcgcgag ttggttcagc tgctgcctga ggctggacga cctcgcggag
     3361 ttctaccggc agtgcaaatc cgtcggcatc caggaaacca gcagcggcta tccgcgcatc
     3421 catgcccccg aactgcagga gtggggaggc acgatggccg ctttggtcga cccggacggg
     3481 acgctcctgc gcctgataca gaacgaattg cttgcaggca tctcatgagt gtgtcttccc
     3541 gttttccgcc tgaggtcact gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc
     3601 tgctcaccac ccactcgcca agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3661 ataggctccg ccgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3721 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3901 tcatgtctgg atcaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     3961 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4021 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4141 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4201 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4261 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4561 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4621 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4681 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4741 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4801 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4861 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4921 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4981 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5041 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5101 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     6121 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6181 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6241 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6301 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6361 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6421 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6481 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6541 atattaacgc ttacaattt
//
