LOCUS       V044846                 7351 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044846
VERSION     V044846
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..2841
                     /label="MAP3K3(NM_002401)"
                     /note="MAP3K3(NM_002401)"
                     /gene="MAP3K3"
     CDS             2848..2913
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2951..3175
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3221..3649
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3663..3992
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4059..4853
                     /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    5027..5148
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5197..5213)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5221..5237
                     /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(5245..5275)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5290..5311
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5599..6184)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6355..7215)
                     /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(7216..7320)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggacg aacaggaggc attgaactca atcatgaacg atctggtggc cctccagatg
     1021 aaccgacgtc accggatgcc tggatatgag accatgaaga acaaagacac aggtcactca
     1081 aataggcaga gtgacgtcag aatcaagttc gagcacaacg gggagaggcg aattatagcg
     1141 ttcagccggc ctgtgaaata tgaagatgtg gagcacaagg tgacaacagt atttggacaa
     1201 cctcttgatc tacattacat gaacaatgag ctctccatcc tgctgaaaaa ccaagatgat
     1261 cttgataaag caattgacat tttagataga agctcaagca tgaaaagcct taggatattg
     1321 ctgttgtccc aggacagaaa ccataacagt tcctctcccc actctggggt gtccagacag
     1381 gtgcggatca aggcttccca gtccgcaggg gatataaata ctatctacca gccccccgag
     1441 cccagaagca ggcacctctc tgtcagctcc cagaaccctg gccgaagctc acctccccct
     1501 ggctatgttc ctgagcggca gcagcacatt gcccggcagg ggtcctacac cagcatcaac
     1561 agtgaggggg agttcatccc agagaccagc gagcagtgca tgctggatcc cctgagcagt
     1621 gcagaaaatt ccttgtctgg aagctgccaa tccttggaca ggtcagcaga cagcccatcc
     1681 ttccggaaat cacgaatgtc ccgtgcccag agcttccctg acaacagaca ggaatactca
     1741 gatcgggaaa ctcagcttta tgacaaaggg gtcaaaggtg gaacctaccc ccggcgctac
     1801 cacgtgtctg tgcaccacaa ggactacagt gatggcagaa gaacatttcc ccgaatacgg
     1861 cgtcatcaag gcaacttgtt caccctggtg ccctccagcc gctccctgag cacaaatggc
     1921 gagaacatgg gtctggctgt gcaatacctg gacccccgtg ggcgcctgcg gagtgcggac
     1981 agcgagaatg ccctctctgt gcaggagagg aatgtgccaa ccaagtctcc cagtgccccc
     2041 atcaactggc gccggggaaa gctcctgggc cagggtgcct tcggcagggt ctatttgtgc
     2101 tatgacgtgg acacgggacg tgaacttgct tccaagcagg tccaatttga tccagacagt
     2161 cctgagacaa gcaaggaggt gagtgctctg gagtgcgaga tccagttgct aaagaacttg
     2221 cagcatgagc gcatcgtgca gtactatggc tgtctgcggg accgcgctga gaagaccctg
     2281 accatcttca tggagtacat gccagggggc tcggtgaaag accagttgaa ggcttacggt
     2341 gctctgacag agagcgtgac ccgaaagtac acgcggcaga tcctggaggg catgtcctac
     2401 ctgcacagca acatgattgt tcaccgggac attaagggag ccaacatcct ccgagactct
     2461 gctgggaatg taaagctggg ggactttggg gccagcaaac gcctgcagac gatctgtatg
     2521 tcggggacgg gcatgcgctc cgtcactggc acaccctact ggatgagccc tgaggtgatc
     2581 agcggcgagg gctatggaag gaaagcagac gtgtggagcc tgggctgcac tgtggtggag
     2641 atgctgacag agaaaccacc gtgggcagag tatgaagcta tggccgccat cttcaagatt
     2701 gccacccagc ccaccaatcc tcagctgccc tcccacatct ctgaacatgg ccgggacttc
     2761 ctgaggcgca tttttgtgga ggctcgccag agaccttcag ctgaggagct gctcacacac
     2821 cactttgcac agctcatgta cctcgaggac tacaaagacc atgacggtga ttataaagat
     2881 catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta aacccgctga
     2941 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3001 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3061 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3121 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3181 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3481 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3541 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3601 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3661 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3721 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3781 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3841 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3901 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3961 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4021 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4081 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4141 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4201 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4261 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4321 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4381 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4441 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4501 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4561 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4621 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4681 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4741 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4801 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4861 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4921 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4981 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5041 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5101 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5161 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5221 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5281 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5341 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5401 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5461 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5521 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5581 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5641 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5701 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5761 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5821 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5881 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5941 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6001 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6061 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6121 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6481 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6961 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7201 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7261 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7321 cacatttccc cgaaaagtgc cacctgacgt c
//
