LOCUS       V040821                 7464 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040821
VERSION     V040821
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..2144
                     /label="Hnrnpk(NM_001360490)"
                     /note="Hnrnpk(NM_001360490)"
                     /gene="Hnrnpk"
     CDS             2151..2216
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2240..2828
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2894..2942
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3144..3572
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3586..3915
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3982..4776
                     /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    4952..5073
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5122..5138)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5146..5162
                     /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(5170..5200)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5215..5236
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5524..6112)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6283..7143)
                     /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(7144..7248)
                     /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 ccaccatgga gaccgaacag ccagaagaaa
      781 ccttccccaa caccgaaacc aatggtgaat ttggtaaacg ccctgcagaa gatatggaag
      841 aggagcaagc ctttaaaaga tctagaaata ctgatgagat ggttgaattg cgcattttgc
      901 ttcagagcaa gaatgctgga gcagtgattg gaaaaggagg caagaatatt aaggctctcc
      961 gtacagacta caatgccagt gtttcagtcc cagacagcag tggccccgag cgcatactga
     1021 gtatcagtgc tgatattgag acgattggag aaattctgaa gaaaatcatc cctaccttgg
     1081 aagagggcct gcagttgcca tcacccactg caaccagcca gctcccgctc gaatctgatg
     1141 ctgtggaatg cttaaattac caacattata aaggaagtga ctttgattgc gagttgagac
     1201 tgttgattca tcagagtctg gcaggaggaa taattggtgt taaaggtgct aaaatcaaag
     1261 aacttcgaga aaacactcag acaacaatca agcttttcca ggagtgctgc cctcactcta
     1321 ctgacagagt tgttcttatt ggaggaaaac ctgatagggt tgtagaatgc atcaagatca
     1381 tccttgacct tatatctgag tctcccatca aaggacgtgc acaaccttat gatcccaact
     1441 tttatgatga gacctatgat tatggtggtt ttacaatgat gtttgatgac cgccgaggac
     1501 gacctgtggg attccccatg aggggaagag gtggttttga cagaatgcct cctggtcggg
     1561 gtgggcgtcc catgcctcct tctagaagag attatgatga tatgagccct cgtcgaggac
     1621 ctccaccacc accacctggt cgaggtggcc ggggtggcag cagagcccgg aatctgcctc
     1681 ttcctcctcc accaccaccc agagggggag atctaatggc ttatgacaga agaggaaggc
     1741 ctggagaccg ctatgatggc atggttgggt tcagtgctga tgaaacttgg gattctgcaa
     1801 ttgacacatg gagcccatca gaatggcaaa tggcttatga accacagggt ggttctggat
     1861 atgactattc ttatgcaggg ggccgtggct catatggtga tcttggcgga cctattatca
     1921 ctacacaagt aactattccc aaagatttgg ctggatctat tattggcaaa ggtggtcagc
     1981 ggattaaaca aattcgtcat gaatctggag catcaatcaa aattgatgaa cctttagaag
     2041 gatctgaaga tcggatcatt accattacag gaacacagga ccagatacag aacgcacagt
     2101 atttgctgca gaacagtgtg aagcagtatt ctggaaagtt tttcggatcc gactacaaag
     2161 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     2221 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2281 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2341 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2401 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2461 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2521 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2581 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2641 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2701 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2761 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2821 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     2881 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     2941 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3001 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3061 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     3121 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3181 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3241 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3301 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3361 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3421 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3481 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     3541 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3661 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3721 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3781 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3841 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3901 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3961 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4021 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4081 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4141 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4201 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4261 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4321 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4381 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4441 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4501 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4561 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4621 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4681 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4741 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     4801 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4861 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4921 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4981 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5041 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5101 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5161 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5221 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5281 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5341 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5401 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5461 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5521 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5581 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5641 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5701 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5761 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5821 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5881 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5941 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6001 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6061 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6121 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6181 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6241 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6301 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6361 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6421 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6481 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6541 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6601 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6661 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6721 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6781 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6841 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6901 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6961 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7021 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7081 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7141 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7201 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7261 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     7321 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7381 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     7441 aattgcatga agaatctgct tagg
//
