LOCUS       V044337                 9939 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044337
VERSION     V044337
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..4619
                     /label="Map4k4(NM_001252200)"
                     /note="Map4k4(NM_001252200)"
                     /gene="Map4k4"
     CDS             4626..4691
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4715..5303
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5369..5417
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5619..6047
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6061..6390
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6457..7251
                     /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    7427..7548
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7597..7613)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7621..7637
                     /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(7645..7675)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7690..7711
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7999..8587)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8758..9618)
                     /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(9619..9723)
                     /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 gaacgactct cccgcgaaga
      781 gcctggtgga cattgacctg tcgtccctgc gggaccctgc tgggattttt gagctggtgg
      841 aagtggttgg aaatggcacc tatggacaag tctataaggg tcgacatgtt aaaacgggtc
      901 agctggccgc catcaaggtt atggacgtca ccgaggatga agaggaagaa atcaaactgg
      961 agataaatat gctgaagaag tattctcatc atcgaaatat tgccacgtac tatggtgctt
     1021 tcattaagaa gagccctcca ggacatgatg accaactctg gcttgttatg gagttttgtg
     1081 gggctgggtc catcacagac cttgtgaaga acaccaaagg gaacactctc aaagaagact
     1141 ggattgctta catctccagg gaaatcctca ggggattggc acatctccat attcaccacg
     1201 ttattcaccg agatatcaag ggccaaaatg tgctgctgac cgagaatgct gaggtgaaac
     1261 ttgttgattt tggtgtaagc gctcagctgg acaggacggt tggacggaga aatacgttca
     1321 taggcacacc ctactggatg gctccagagg tcatcgcctg tgatgagaac ccagacgcca
     1381 cttacgacta cagaagtgac ctctggtcct gtggcatcac agccatcgag atggctgaag
     1441 gggccccccc tctctgtgac atgcatccaa tgagagcgct gtttctcatc cccagaaacc
     1501 ctcctcccag gctgaagtca aaaaaatggt caaagaaatt tttcagcttt atagaaggct
     1561 gtctggtgaa gaattacatg cagcggccct ctacagagca acttttaaaa caccctttca
     1621 taagggatca gcccaatgaa aggcaggttc gaatccagct taaggatcac atagaccgga
     1681 ccagaaagaa gagaggcgag aaagatgaga cggagtacga gtacagcggg agcgaggagg
     1741 aggaggagga agtgcctgag caggagggag agccaagttc catcgtcaat gtgcctggag
     1801 agtcaactct gcgacgtgat ttcctgagac tgcagcagga gaacaaggag cggtctgagg
     1861 ctctgcggag acagcagctt ctgcaggagc agcagctccg ggagcaggag gagtataaga
     1921 ggcagctgct ggctgagagg cagaagcgga ttgaacagca gaaagaacag aggaggcggc
     1981 tggaagagca acaaagaaga gaacgggaag ccaggaggca gcaggagcgt gagcagcggc
     2041 ggcgtgaaca agaggagaag aggcgtctcg aggaactgga aaggcggcgt aaagaagagg
     2101 aagagaggag acgggcagaa gaggagaaga ggagagtgga gagggaacag gagtacatca
     2161 ggcggcagct agaggaggag cagcggcacc tggagatcct gcagcagcag ctgctccagg
     2221 agcaggccat gttactgcac gaccacagga ggccgcacgc acagcagcag ccgccgcccc
     2281 cgcagcagca ggacaggagc aaaccgagct ttcatgctcc agagcccaag cctcactatg
     2341 accctgctga cagagctcgg gaggtggaag atagatttag aaagactaac cacagctccc
     2401 ctgaggccca ggctaagcaa acgggcagag gactggagcc acccgtgcct tcccggtcag
     2461 agtctttttc caatggcaac tccgagtctg tgcaccctgc cctgcagaga ccagcggagc
     2521 cacaggtaca gtggtcccac ctggcatctc tcaagaacaa tgtctcccct gtctcgagat
     2581 cccattcctt cagtgaccct tctcccaaat tcgcacacca ccatctccgc tctcaggacc
     2641 catgtccacc ttcccgcagt gaggggctca gtcagagctc tgactctaag tcggaggtgc
     2701 ccgagcccac ccaaaaggct tggtctagat cagacagtga cgaggtgcct ccaagggttc
     2761 ccgtgagaac gacctctcgt tcccctgtcc tctcccgtcg ggattcccca ctgcagggcg
     2821 gtggacagca gaatagccaa gcaggacaga gaaattccac cagcagtatt gagccccggc
     2881 ttctttggga gagagtggaa aagctggtcc ccaggccagg cagtggcagc tcttcagggt
     2941 ccagcaactc aggatcccag cctggctccc atcctgggtc tcagagcggc tccggtgaac
     3001 gcttcagagt gagatcatca tccaagtctg aaggttctcc atcgccacgc caggaaagtg
     3061 cagccaaaaa gcctgacgat aagaaagaag tattccggcc tctcaagcct gcgggagaag
     3121 tggacttgac tgcgttggcc aaagagcttc gagcagtgga agatgttcgg ccacctcaca
     3181 aagtaacaga ctactcatcc tccagtgagg agtctgggac cacagatgag gaggaggagg
     3241 acgtggaaca ggagggggct gatgattcta cctcgggacc agaagacacc agagcagcgt
     3301 catctccgaa cctgagcaac ggtgaaacag aatctgtgaa aacaatgatc gttcatgatg
     3361 atgtagaaag tgagccagcc atgaccccgt ccaaggaggg caccctcatc gtccgccaga
     3421 ctcagtccgc tagtagcaca ctccagaaac acaaatcttc ctcctccttt acacctttta
     3481 tagaccccag gttactacag atctctccat ctagtgggac gacagtgact tccgtggtgg
     3541 gattttcctg tgatgggctg agaccagaag ccataaggca agatcctact cggaaaggct
     3601 cagtggtcaa tgtgaacccc acgaacacta ggccacagag tgataccccc gagattcgta
     3661 aatacaagaa gagatttaac tcagagatcc tgtgtgctgc cttatgggga gtgaacttgc
     3721 tggttggtac agagagcggt ctgatgctct tggacagaag tggccaaggg aaagtatacc
     3781 ctctgatcag ccgaagacgg ttccagcaaa tggatgtgct cgaaggccta aatgtcctgg
     3841 tgacaatatc tggcaaaaag gataagttac gtgtctacta tttatcctgg ctaagaaata
     3901 agattcttca taatgatcca gaggttgaga agaagcaggg gtggaccact gtgggcgact
     3961 tggaagggtg cgtgcactat aaagtcgtaa aatatgaaag aatcaagttt ctggtaattg
     4021 ctttgaagag ttctgtggaa gtctatgcat gggcaccgaa gccatatcac aaatttatgg
     4081 cctttaagtc atttggagaa ctgctacata agccattact ggtggatctc actgtggagg
     4141 aaggccagag gttgaaagtg atctatggat cctgtgcggg attccatgct gttgatgtgg
     4201 attcaggctc agtctatgac atttatctac caacacatat tcagtgtagc atcaaacccc
     4261 atgcaatcat tatcctcccc aacacggatg ggatggagct gctggtgtgc tacgaagacg
     4321 agggggttta cgttaacacg tacggaagga tcaccaagga cgtggttctg cagtggggag
     4381 agatgccgac atctgtagca tatatccggt cgaatcagac aatgggctgg ggagagaagg
     4441 caatagagat acgatctgtg gaaactggtc acttggatgg tgtatttatg cacaaaaggg
     4501 ctcagagact aaagttcctg tgtgaacgca atgacaaggt gttctttgcc tccgtccgct
     4561 ctggtggcag cagccaggtc tatttcatga ccttaggcag gacttctctt ctgagctggg
     4621 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4681 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     4741 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4801 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4861 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4921 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4981 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5041 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5101 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5161 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5221 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5281 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     5341 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     5401 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5461 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5521 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5581 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     5641 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5701 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5761 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5821 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5881 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5941 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     6001 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     6061 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6121 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6181 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6241 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6301 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     6361 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     6421 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6481 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6541 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6601 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6661 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6721 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6781 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6841 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6901 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6961 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7021 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7081 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     7141 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     7201 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     7261 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7561 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8161 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     8221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8401 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9721 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9781 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9841 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     9901 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
