LOCUS       V019654                 9356 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019654
VERSION     V019654
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"
     CDS             683..4036
                     /label="MAP3K9(NM_033141)"
                     /note="MAP3K9(NM_033141)"
                     /gene="MAP3K9"
     CDS             4043..4108
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4132..4720
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4786..4834
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5036..5464
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5478..5807
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5874..6668
                     /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    6844..6965
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7014..7030)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7038..7054
                     /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(7062..7092)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7107..7128
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7416..8004)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8175..9035)
                     /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(9036..9140)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggagcc ctccagagcg cttctcggct gcctagcgag
      721 cgccgccgct gccgccccgc cgggggagga tggagcaggg gccggggccg aggaggagga
      781 ggaggaggag gaggaggcgg cggcggcggt gggccccggg gagctgggct gcgacgcgcc
      841 gctgccctac tggacggccg tgttcgagta cgaggcggcg ggcgaggacg agctgaccct
      901 gcggctgggc gacgtggtgg aggtgctgtc caaggactcg caggtgtccg gcgacgaggg
      961 ctggtggacc gggcagctga accagcgggt gggcatcttc cccagcaact acgtgacccc
     1021 gcgcagcgcc ttctccagcc gctgccagcc cggcggcgag gaccccagtt gctacccgcc
     1081 cattcagttg ttagaaattg attttgcgga gctcaccttg gaagagatta ttggcatcgg
     1141 gggctttggg aaggtctatc gtgctttctg gataggggat gaggttgctg tgaaagcagc
     1201 tcgccacgac cctgatgagg acatcagcca gaccatagag aatgttcgcc aagaggccaa
     1261 gctcttcgcc atgctgaagc accccaacat cattgcccta agaggggtat gtctgaagga
     1321 gcccaacctc tgcttggtca tggagtttgc tcgtggagga cctttgaata gagtgttatc
     1381 tgggaaaagg attcccccag acatcctggt gaattgggct gtgcagattg ccagagggat
     1441 gaactactta catgatgagg caattgttcc catcatccac cgcgacctta agtccagcaa
     1501 catattgatc ctccagaagg tggagaatgg agacctgagc aacaagattc tgaagatcac
     1561 tgattttggc ctggctcggg aatggcaccg aaccaccaag atgagtgcgg cagggacgta
     1621 tgcttggatg gcacccgaag tcatccgggc ctccatgttt tccaaaggca gtgatgtgtg
     1681 gagctatggg gtgctacttt gggagttgct gactggtgag gtgccctttc gaggcattga
     1741 tggcttagca gtcgcttatg gagtggccat gaacaaactc gcccttccta ttccttctac
     1801 gtgcccagaa ccttttgcca aactcatgga agactgctgg aatcctgatc cccactcacg
     1861 accatctttc acgaatatcc tggaccagct aaccaccata gaggagtctg gtttctttga
     1921 aatgcccaag gactccttcc actgcctgca ggacaactgg aaacacgaga ttcaggagat
     1981 gtttgaccaa ctcagggcca aagaaaagga acttcgcacc tgggaggagg agctgacgcg
     2041 ggctgcactg cagcagaaga accaggagga actgctgcgg cgtcgggagc aggagctggc
     2101 cgagcgggag attgacatcc tggaacggga gctcaacatc atcatccacc agctgtgcca
     2161 ggagaagccc cgggtgaaga aacgcaaggg caagttcagg aagagccggc tgaagctcaa
     2221 ggatggcaac cgcatcagcc tcccttctga tttccagcac aagttcacgg tgcaggcctc
     2281 ccctaccatg gataaaagga agagtcttat caacagccgc tccagtcctc ctgcaagccc
     2341 caccatcatt cctcgccttc gagccatcca gttgacacca ggtgaaagca gcaaaacctg
     2401 gggcaggagc tcagtcgtcc caaaggagga aggggaggag gaggagaaga gggccccaaa
     2461 gaagaaggga cggacgtggg ggccagggac gcttggtcag aaggagcttg cctcgggaga
     2521 tgaaggatcc cctcagagac gtgagaaagc taatggttta agtaccccat cagaatctcc
     2581 acatttccac ttgggcctca agtccctggt agatggatat aagcagtggt cgtccagtgc
     2641 ccccaacctg gtgaagggcc caaggagtag cccggccctg ccagggttca ccagccttat
     2701 ggagatggcc ttgctggcag ccagttgggt ggtgcccatc gacattgaag aggatgagga
     2761 cagtgaaggc ccagggagtg gagagagtcg cctacagcat tcacccagcc agtcctacct
     2821 ctgtatccca ttccctcgtg gagaggatgg cgatggcccc tccagtgatg gaatccatga
     2881 ggagcccacc ccagtcaact cggccacgag tacccctcag ctgacgccaa ccaacagcct
     2941 caagcggggc ggtgcccacc accgccgctg cgaggtggct ctgctcggct gtggggctgt
     3001 tctggcagcc acaggcctag ggtttgactt gctggaagct ggcaagtgcc agctgcttcc
     3061 cctggaggag cctgagccac cagcccggga ggagaagaaa agacgggagg gtctttttca
     3121 gaggtccagc cgtcctcgtc ggagcaccag ccccccatcc cgaaagcttt tcaagaagga
     3181 ggagcccatg ctgttgctag gagacccctc tgcctccctg acgctgctct ccctctcctc
     3241 catctccgag tgcaactcca cacgctccct gctgcgctcc gacagcgatg aaattgtcgt
     3301 gtatgagatg ccagtcagcc cagtcgaggc ccctcccctg agtccatgta cccacaaccc
     3361 cctggtcaat gtccgagtag agcgcttcaa acgagatcct aaccaatctc tgactcccac
     3421 ccatgtcacc ctcaccaccc cctcgcagcc cagcagtcac cggcggactc cttctgatgg
     3481 ggcccttaag ccagagactc tcctagccag caggagcccc tccagcaatg ggttgagccc
     3541 cagtcctgga gcaggaatgt tgaaaacccc cagtcccagc cgagacccag gtgaattccc
     3601 ccgtctccct gaccccaatg tggtcttccc cccaacccca aggcgctgga acactcagca
     3661 ggactctacc ttggagagac ccaagactct ggagtttctg cctcggccgc gtccttctgc
     3721 caaccggcaa cggctggacc cttggtggtt tgtgtccccc agccatgccc gcagcacctc
     3781 cccagccaac agctccagca cagagacgcc cagcaacctg gactcctgct ttgctagcag
     3841 tagcagcact gtagaggagc ggcctggact tccagccctg ctcccgttcc aggcagggcc
     3901 gctgcccccg actgagcgga cgctcctgga cctggatgca gaggggcaga gtcaggacag
     3961 caccgtgccg ctgtgcagag cggaactgaa cacacacagg cctgcccctt atgagatcca
     4021 gcaggagttc tggtctctcg aggactacaa agaccatgac ggtgattata aagatcatga
     4081 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     4141 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4201 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4261 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4321 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4381 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     4441 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4501 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     4561 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4621 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4681 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     4741 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4801 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4861 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4921 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4981 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     5041 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5101 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5161 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     5221 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5281 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5341 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5401 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5461 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5521 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5581 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5641 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5701 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     5761 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     5821 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5881 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5941 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6001 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6061 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6121 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6181 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6241 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6301 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6361 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     6421 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6481 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6541 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6601 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6661 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     6721 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6781 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6841 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6901 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6961 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     7021 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7081 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7141 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7201 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     7621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     9181 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     9241 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     9301 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
