LOCUS       V038918                 8454 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038918
VERSION     V038918
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..3134
                     /label="Ufl1(NM_026194)"
                     /note="Ufl1(NM_026194)"
                     /gene="Ufl1"
     CDS             3141..3206
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3230..3818
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3884..3932
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4134..4562
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4576..4905
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4972..5766
                     /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    5942..6063
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6112..6128)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6136..6152
                     /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(6160..6190)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6205..6226
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6514..7102)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7273..8133)
                     /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(8134..8238)
                     /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 ccaccatggc ggacgcctgg gaggagatcc
      781 ggcggctggc ggctgacttc cagcgggcgc agtttgcaga gtccacgcaa aggttgtcag
      841 agcggaactg cattgagatt gtgaataaat tgatctctca gaaacagctg gaagtagtcc
      901 atacacttga cggcaaggaa tatatcactc cagcccaaat cagtaaagaa atgagagacg
      961 agctacatgt ccgaggtggt cgagtaaaca ttgttgactt acaacaggtt attaatgtgg
     1021 acctgactca tatcgagagt agagtcagtg atattattaa atcagagaag cacgtgcaga
     1081 tggtactggg acaactgatt gatgagaact atctggatca gttatcagaa gaggtgaatg
     1141 ataaattgca agaaagtggt caagtaactg tctcagaact gtgtaaagcc tatgaccttc
     1201 ctggggattt tctgacacag gcattaactc agcgacttgg ccggattatc aatggccatc
     1261 tggaccttga taacagagga gtgattttca ctgaggcctt tgtagctcgc cataaagcac
     1321 gtatccgcgg actcttcagt gccattacac ggcctacacc agtgaactct ttggtttcaa
     1381 aatatggatt tcaggaacag ctcctttact ctgtgctcga ggacctcgtc agcactggcc
     1441 gcttacgagg caccgtggtc ggcgggcgac aggataaagc agtctttgtc cctgacatct
     1501 actccaggac acagagtact tgggtggact cctttttcag gcagaatggc tatctagaat
     1561 ttgacgcttt atccagactt ggaatcccag atgctgtgaa ctatataaag aaaaggtata
     1621 agaataccca actcttgttt ttgaaggcaa cttgtgttgg ccaaggactc gtagaccagg
     1681 tggaagcatc cgtagaggaa gccatcagct cagggacctg ggttgacatt tcgcctctgc
     1741 tacccagttc cttgtcagtt gaagatgctg ctatgctgct ccagcaggtg atgagaccat
     1801 tcggcaagct cgcttcagct atagtcttta gtgacacggt tgtggtgagt gaaaagttca
     1861 taactgactg cacaggactg ttcagcgaac ggatgcacca aaaagctgag aaggaaatga
     1921 aaaataatcc tgtgcattta atcactgaag aagatctgaa acaaatttcc attttagaaa
     1981 gtgttaatac aagtaaaaag gataaaaaag atgaacgcag gaagaaagca acagagggct
     2041 ctggaagtgt aagaggagga ggagggggca atgccagaga atataaaatt aagaaaacca
     2101 agaagaaggg aagaaaagat gaggacagtg atgatgagtc ccagtcgtcg catggtggaa
     2161 agaagaagcc ggacatcaca ttcatgttcc aggatgaaat tgaagattgc ttaaggaaac
     2221 acattcagga tgcccctgag gagtttattt cagagcttgc agaatactta ataaagcctc
     2281 ttaataaaat gtatcttgag gtagtacgtt cagtcttcat gtcttcaaca tctgcttctg
     2341 gaactggcag aaaacgcaca atcaaggact tacaagaaga ggtttcaaac ctatacaata
     2401 atatcagatt atttgaaaag ggaatgaaat attttgcaga tgacacacag actgctctta
     2461 ccaagcactt gctgaagaca gtgtgtaccg atatcactaa cctcatgttc aacttcctag
     2521 cctcagattt cctcatggca gtggaggagc ctgcagccat tacaagtgat ataagaaaaa
     2581 aaattttaag caaattgaca gaagaaacca aagtagctct cacaaaactc cataactctc
     2641 tcaatgaaaa aagcatagaa gacttcctgt cttgcctgga ttctgccaca gaagcgtgtg
     2701 acattatggt gaaaaaggga gacaagaaaa gggaaagaca aatcctcttc cagcatcgac
     2761 aggctctgtg tgagcagctg aaggtcacag aagaccctgc tctgatttta cacctcacag
     2821 cagtcctgtt atttcagctc tcaacccaca gcatgctcca cgcacctgga aggtgtgtcc
     2881 cacagatcat cgcgtttctt cacagcaaaa tcccagagga tcaacatact ctcttggtaa
     2941 aatatcaagg tttggttgtg aaacagttag tcagtcaaaa taagaagact gggcagggag
     3001 aggatccgtc aagcgatgaa ttagacaagg aacaacatga tgtcacaaac gctactcgga
     3061 aagagcttca ggaactgtca ttatccatta aggaccttgt tctcaagtcc aggaaatcat
     3121 ctgtcacaga ggagggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     3181 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3241 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3301 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3361 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3421 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3481 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3541 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3601 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3661 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3721 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3781 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3841 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3901 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3961 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4021 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4081 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     4141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4201 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4261 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4321 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4441 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4501 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4561 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4621 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4681 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4741 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4801 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4861 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4921 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4981 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5041 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5101 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5161 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5221 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5281 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5341 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5401 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5461 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5521 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5581 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5641 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5701 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5761 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5821 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5881 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5941 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6001 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6061 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6121 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6181 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6241 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6301 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6361 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6421 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7081 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7201 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7261 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7321 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7381 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7441 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7501 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7561 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7621 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7681 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7741 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7801 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7861 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7921 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7981 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8041 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8101 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8161 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8221 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8281 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8341 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8401 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
