LOCUS       V028404                 9330 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028404
VERSION     V028404
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4010
                     /label="Ube4a(NM_001346698)"
                     /note="Ube4a(NM_001346698)"
                     /gene="Ube4a"
     CDS             4017..4082
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4106..4694
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4760..4808
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5010..5438
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5452..5781
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5848..6642
                     /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    6818..6939
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6988..7004)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7012..7028
                     /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(7036..7066)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7081..7102
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7390..7978)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8149..9009)
                     /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(9010..9114)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgac agaccaggag aataacaaca
      781 acatctcaag taaccccttt gctgctcttt ttggctccct ggctgatgcc aagcagtttg
      841 cagcaatcca gaaagagcag ctgaagcagc agtccggtag tgtatttggt tctgaggaag
      901 aagcgttgtg taacagctta catgttccag cagatgagct cccagcaagc ccagatgact
      961 cggataatag tgtgtcagag agcctggatg agtttgacta ctctgtatct gagattagcc
     1021 gctccttccg gacacaccaa gaaatgtgtg agcaactcaa catcaatcac atgatccaga
     1081 ggatcttcct catcaccctg gacaacagtg atccaagctt gaaaagcggg aatggcatcc
     1141 ccagccgttg tgtgtacttg gaagaaatgg cagtagagct ggaagatcaa gactggcttg
     1201 atatgagcaa tgttgagcag gccatctttg ctcgattatt gcttcaagat ccaggaaacc
     1261 acttgatcag catgacttcc tccacaactt taaatctctc tgctgatcgt gatgcaggag
     1321 agaggcacat cttctgttac ctttactcct gcttccagag agccaaggaa gagattacca
     1381 aagttccaga aaacctactc ccctttgcag tgcagtgcag aaacctcact gtgtccaaca
     1441 ctcgaacagt tctcctcacc ccagagatct atgttgacca aaacatccat gagcaactgg
     1501 tagatttgat gttggaagct attcagggag cccattttga agatgtaact gaatttttag
     1561 aagaggtcat tgaagccttg ctgttggatg aggaggttcg aacatttcct gaagtcatga
     1621 ttccagtgtt tgatatttta ctcagccgaa taaaagatct ggaactctgc cagatcctcc
     1681 tgtatgcata tctggacatt cttctctatt tcactaggca gaaggatatg gccaaggtgt
     1741 ttttagaata cattcagccc aaggatccca gcaatgggca gatgtatcag aagaccttgc
     1801 tgggagtcat cctgaatatc tcgtgcttgt taaagactcc gggtgtggta gagaaccatg
     1861 gcttcttcct gaatccatcc cgttccagtc ctcaggagat caaagtacag gaggccaaca
     1921 tccatcagtt catggctcag ttccatgaaa agatctacca gatgctgaag aacttgctcc
     1981 agctctctcc agaaaccaag cactgtatct tgttctggct tggaaactgt ttgcatgcaa
     2041 atgcaggccg caccaagatc tgggccaatc agatgcccga aatcttcttc caaatgtatg
     2101 cctcggatgc cttctttctg aatctgggtg ctgccctcct gaagctgtgc cagccattct
     2161 gcaagcccag atcctcccgg ctcctcacct ttaaccccac gtactgtgtc ctcaaggact
     2221 tgaatgatga agaacgaaaa attaaaagtg tgcatatgag aggtttggac aaggagacct
     2281 gtctgatccc agctgtgcag gagccaacgt tcccccagag ctacaacctt gtgacggaga
     2341 acctggccct gacagagtat accttgtact tgggatttca caggttgcat gatcagatgg
     2401 taaaaatcaa ccaaaatctg catcggctgc aggttgcctg gcgggatgct cagcagagtt
     2461 ctagccctgc tgctgacaat ctccgtgagc agttcgaacg cctgatgacc atctatctct
     2521 ctaccaagac tgctatgact gaaccccaga tgctgcaaaa ctgtctgaat ttgcaggtgt
     2581 ccatggctgt tctgctggtt caactggcca taggcaatga gggctcccag ccaatagaac
     2641 taagtttccc tttgccagat ggctacagct ctttggctta tgtgccagaa ttttttgcag
     2701 ataacttggg tgatttcctc attttcctcc ggcgctttgc tgaggacatc ttggagacgt
     2761 cagcagactc cctggagcat gtccttcatt ttatcaccat tttcactgga agcatagaaa
     2821 gaatgaagaa tcctcacctg agggccaaac tagcagaggt gttagaagca gtgatgcctc
     2881 acctagacca gaccccaagc cccttggtgt ccagtgtgtt ccaccggaaa cgtgtgttct
     2941 gcaactttcc atacgctccc caacttgcag aagctttgat caaggtgttt gtggacatcg
     3001 agtttacagg ggacccccat caatttgaac agaagtttaa ttaccgccgt cctatgtatc
     3061 ccatcctaag atacatgtgg gggacagatt gctatcggga gagcattaag gatttggctg
     3121 actatgcctc taagaattta gaagctatga atcccccact gttcctccga tttcttaacc
     3181 tgttaatgaa tgatgctatc ttccttttgg atgaagccat acagtatttg agcaagataa
     3241 agattcagca gattgagaaa gaccgaggag aatgggagag tctgacacca gaagcccgcc
     3301 gagagaaaga ggcaggtctg cagatgtttg ggcagctggc acggttccat aacatcatgt
     3361 ccaatgaaac aattggtacc ctttccttcc tgacatcaga gatcaagtcg ctgtttgtgc
     3421 atccgttcct ggccgagcgc atcatctcca tgctcaacta cttcctgcag cacctggtgg
     3481 gccccaagat gggtgccctc aaagtcaagg acttcagcga atttgacttc aaacctcagc
     3541 agctcgtatc agacatctgc actatctact taaatcttgg ggatgaggag aacttctgtg
     3601 ccactgtgcc caaagacgga cgttcctatt ctccgactct ctttgcacag acagtccgtg
     3661 tcctgaagaa aatcaataag cccgggaata tgatcgtggc tttcagtaac ttggcagaga
     3721 gaatcaagtc tcttgcagac ctccaacaac aagaggagga gacatatgca gacgcctgtg
     3781 atgagttcct ggatcccatc atgagcacac tgatgtctga ccctgtggtg cttccatctt
     3841 ccagagtcac tgtggacaga tccaccattg ctaggcattt actcagtgac caaacagatc
     3901 cttttaaccg tagtcccctt accatggacc agatccggcc caacacagaa ctgaaagaaa
     3961 aaatccaacg gtggctcgca gaaaggaagc aacaaaagga gcagcctgaa ggatccgact
     4021 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     4081 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     4141 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4201 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4261 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4321 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4381 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4441 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4501 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4561 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4621 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4681 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     4741 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     4801 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     4861 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     4921 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     4981 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     5041 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5101 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5161 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5221 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5281 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5341 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     5401 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     5461 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5521 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5581 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5641 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5701 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     5761 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     5821 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5881 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5941 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     6001 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     6061 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     6121 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     6181 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     6241 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     6301 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     6361 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6421 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6481 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6541 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6601 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6661 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6721 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6781 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6841 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6901 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6961 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7021 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7081 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7141 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7201 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7561 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     7621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7801 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8161 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8221 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8281 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8341 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8401 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8461 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8521 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8581 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8641 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8701 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8761 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8821 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8881 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8941 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9001 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9061 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9121 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     9181 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     9241 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     9301 accgacaatt gcatgaagaa tctgcttagg
//
