LOCUS       V035678                 8305 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035678
VERSION     V035678
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        431..708
                     /label="chicken β-actin promoter"
     intron          709..1726
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     CDS             1789..2985
                     /label="MAP2K4(NM_003010)"
                     /note="MAP2K4(NM_003010)"
                     /gene="MAP2K4"
     CDS             2992..3057
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3081..3669
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3735..3783
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3985..4413
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4427..4756
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4823..5617
                     /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    5793..5914
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5963..5979)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5987..6003
                     /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(6011..6041)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6056..6077
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6365..6953)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7124..7984)
                     /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(7985..8089)
                     /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 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      481 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      541 gggggggggg ggcccccccc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      601 cggaaaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      661 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgttgcg
      721 cgctgccttc cccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      781 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      841 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      901 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
      961 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     1021 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1081 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     1141 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     1201 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     1261 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1321 gccggggagg gctcggggga aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
     1381 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
     1441 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     1501 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
     1561 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1621 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1681 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1741 gctggttatt gtgctgtctc atcattttgg caaagaattc tcgccaccat ggcggctccg
     1801 agcccgagcg gcggcggcgg ctccgggggc ggcagcggca gcggcacccc cggccccgta
     1861 gggtccccgg cgccaggcca cccggccgtc agcagcatgc agggtaaacg caaagcactg
     1921 aagttgaatt ttgcaaatcc acctttcaaa tctacagcaa ggtttactct gaatcccaat
     1981 cctacaggag ttcaaaaccc acacatagag agactgagaa cacacagcat tgagtcatca
     2041 ggaaaactga agatctcccc tgaacaacac tgggatttca ctgcagagga cttgaaagac
     2101 cttggagaaa ttggacgagg agcttatggt tctgtcaaca aaatggtcca caaaccaagt
     2161 gggcaaataa tggcagttaa aagaattcgg tcaacagtgg atgaaaaaga acaaaaacaa
     2221 cttcttatgg atttggatgt agtaatgcgg agtagtgatt gcccatacat tgttcagttt
     2281 tatggtgcac tcttcagaga gggtgactgt tggatctgta tggaactcat gtctacctcg
     2341 tttgataagt tttacaaata tgtatatagt gtattagatg atgttattcc agaagaaatt
     2401 ttaggcaaaa tcactttagc aactgtgaaa gcactaaacc acttaaaaga aaacttgaaa
     2461 attattcaca gagatatcaa accttccaat attcttctgg acagaagtgg aaatattaag
     2521 ctctgtgact tcggcatcag tggacagctt gtggactcta ttgccaagac aagagatgct
     2581 ggctgtaggc catacatggc acctgaaaga atagacccaa gcgcatcacg acaaggatat
     2641 gatgtccgct ctgatgtctg gagtttgggg atcacattgt atgagttggc cacaggccga
     2701 tttccttatc caaagtggaa tagtgtattt gatcaactaa cacaagtcgt gaaaggagat
     2761 cctccgcagc tgagtaattc tgaggaaagg gaattctccc cgagtttcat caactttgtc
     2821 aacttgtgcc ttacgaagga tgaatccaaa aggccaaagt ataaagagct tctgaaacat
     2881 ccctttattt tgatgtatga agaacgtgcc gttgaggtcg catgctatgt ttgtaaaatc
     2941 ctggatcaaa tgccagctac tcccagctct cccatgtatg tcgatctcga ggactacaaa
     3001 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     3061 tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3121 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3181 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3241 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3301 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3361 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3421 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3481 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3541 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3601 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3661 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3721 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     3781 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3841 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3901 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     3961 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4021 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4081 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4141 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4201 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4261 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4321 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     4381 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4501 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4561 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4621 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4681 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4741 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4801 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4861 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4921 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4981 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5041 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5101 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5161 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5221 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5281 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5341 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5401 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5461 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5521 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5581 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     5641 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5701 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5761 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5881 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5941 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6001 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6061 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6121 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6181 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6241 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6301 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6361 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6481 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6541 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6601 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6661 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6721 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6781 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6841 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6901 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6961 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7021 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7081 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7141 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7201 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7261 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7321 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7381 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7441 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7501 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7561 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7621 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7681 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7741 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7801 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7861 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7921 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7981 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8041 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8101 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     8161 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     8221 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     8281 caattgcatg aagaatctgc ttagg
//
