LOCUS       V030332                 7919 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030332
VERSION     V030332
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3388
                     /label="Map4k1(NM_008279)"
                     /note="Map4k1(NM_008279)"
                     /gene="Map4k1"
     CDS             3395..3412
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             3416..3439
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    3486..3710
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3756..4184
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4198..4527
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4594..5388
                     /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    5562..5683
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5732..5748)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5756..5772
                     /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(5780..5810)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5825..5846
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6134..6719)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6890..7750)
                     /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(7751..7855)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gcccttgtgg accccgacat ttttaataag gaccctcgag aacactatga
      961 tctactgcaa cgactgggtg gtggaactta tggagaagta ttcaaggctc gcgacaaggt
     1021 gtctaaggac ctggtggctt tgaagatggt gaagatggag cctgatgatg acgtcgccac
     1081 ccttcagaag gagattctca tgctgaaaac ttgccggcac gccaacatcg tggcttacca
     1141 tggcagttac ctctggctgc agaagctgtg gatctgcatg gaattctgtg gagctggttc
     1201 tctccaggac atctaccaag tgactggctc tctgtccgag ctccagatca gctatgtgtg
     1261 ccgggaagtg cttcagggac tggcctacct gcattcagag aagaagatac atcgagacat
     1321 caagggagcc aacatcctca tcaacgactg cggggaggtt aagctggctg actttgggat
     1381 ctcagcccag attggggcga cactggcccg gcgcctttct ttcattggga caccgtactg
     1441 gatggctcca gaggtggcag cggtggccct gaagggtgga tacaatgagc tgtgcgacat
     1501 ctggtctctt ggcatcacag ccatcgagct ggctgagctt cagccccctc tctttgatgt
     1561 gcaccctctc agggttctct tcctcatgac caagagcggg taccagcctc ctcggctgaa
     1621 ggaaaagagc agatggtcat cttccttcca caactttgtc aaagtcaccc tcactaagaa
     1681 ctccaagaaa cgccccagtg ccaccaagat gctcagtcat cagctggtgt cccagccagg
     1741 gctgaacaga ggcctgatct tggatcttct tgacaaaatg aagaatcctg ggaaggggct
     1801 gcctgttgat attgaagatg aggagcctga gccaccccct gccatccctc ggcggatcag
     1861 atctacctac cgggccagct cactagggat cccagatgca gactgctgcc ggaggcaaat
     1921 ggagttccag aggcccagat gcgtggactg cagacctcag gctgaaactg ttcgcttata
     1981 tccccctgca cacttcggga gcaccagtcc caggagtcag ctgtcagatt ctgatgacga
     2041 ctatgatgat gtggacatcc ctgccccttc agagaacata cctcctccac tgccccccaa
     2101 gcccaagttt cggtctccat cagatgatgg ttctggagga attagggacg atgggcaatt
     2161 gagcccaggg gttctggttc gatgtgcaag tgggccccct ccacgtactc cccgacctgg
     2221 acctccccca gcgacctgca gccctcacct cacagctcgc tcagatccct cattgtggaa
     2281 cccagctgct ccggagccag gccagccccc actcgtaccc ccaaggaagg aaaagatgag
     2341 gggaaagatg gagaacgaga agcgcagaga gaaatatccc cttcttgtga aattattcaa
     2401 tggctgtccc cttcagatcc acagcacagc tgcctggaca catccctcta caaaagatca
     2461 gaacctgctc ctgggggcag aggaaggcat tttcatactg aaccggaatg atcaggaggc
     2521 cacgctggaa atgatcttcc ctggccgcac aacctggctc tattgcatca acaacctcct
     2581 catgtctctc tcagggaaga caccctacct gtattctcat agcatcctgg gcctgcttga
     2641 aaggaaagac ggcagaacgg gaagccccat tgctcacatt agccctcacc ggttactggc
     2701 aagaaagaac atggtctcca gtaagatcca ggacaccaaa ggctgccggg cttgctgtgt
     2761 ggctgagagt gctagctctg ggggcccatt cttatgtggg gcattggaga cgtctgtggt
     2821 tttgctccag tggtaccaac caatgaacaa attcttgctt gtccggcagg tgctgttccc
     2881 gctgcccacg ccgctgccgg tgttcacgct gctgacaacg ccgggctccg agctgccagc
     2941 tgtgtgcatc ggcgtgagcc cggggcaggc ggcaaagtcg gtgctcttcc acaccgtgcg
     3001 cttcggtgcg ctgtcctgct ggctggacga ctccagcacg gagcacaagg gaccagtgca
     3061 ggtgatccaa gtaaaggaag acatggtgat ggtgttgatg gatggctctc taaagctagt
     3121 gaccccagag ggggcaccag ctcctgggct ccgcacccct gagatcccca tgacagaagc
     3181 ggtcgaggct gtggctatgg ttgaagatcg actcgaggcc ttctggaagc atggagttca
     3241 ggtgtgggct ccaggcttga aacagccgct gcaggagctg agagacccca ccctcacctt
     3301 ccgtctgctc tgctccccca ggccggtagt ggtggagaca aggcccacgg atgaccctac
     3361 ggcccccagc aacctctaca tccaggaaga attccatcat catcatcatc atgcggatta
     3421 caaggatgac gacgataagt gactcgagtc tagagggccc gtttaaaccc gctgatcagc
     3481 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3541 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3601 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3661 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3721 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3781 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3841 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3901 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3961 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4021 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4081 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4141 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4381 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4441 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4501 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4561 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4621 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4681 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4741 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4801 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4861 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4921 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4981 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5041 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5101 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5161 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5221 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5281 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5341 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5401 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5461 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5521 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5581 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5641 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5701 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5761 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5821 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5881 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5941 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6001 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6061 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6241 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6301 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6361 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6421 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6481 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6541 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6601 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6661 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6721 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6781 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6841 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6901 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6961 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7021 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7081 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7141 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7201 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7261 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7321 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7381 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7441 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7501 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7561 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7621 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7681 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7741 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7801 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7861 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
