LOCUS       V020958                 8282 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020958
VERSION     V020958
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             677..2962
                     /label="KSR1(NM_014238)"
                     /note="KSR1(NM_014238)"
                     /gene="KSR1"
     CDS             2969..3034
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3058..3646
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3712..3760
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3962..4390
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4404..4733
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4800..5594
                     /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    5770..5891
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5940..5956)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5964..5980
                     /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(5988..6018)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6033..6054
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6342..6930)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7101..7961)
                     /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(7962..8066)
                     /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 ggatccatga atgaggccaa ggtgaaggag acgctgcggc gctgtggggc
      721 cagcggggat gagtgtggcc gtctgcagta tgccctcacc tgcctgcgga aggtgacagg
      781 cctgggaggg gagcacaagg aggactccag ttggagttca ttggatgcgc ggcgggaaag
      841 tggctcaggg ccttccacgg acaccctctc agcagccagc ctgccctggc ccccagggag
      901 ctcccagctg ggcagagcag gcaacagcgc ccagggccca cgctccatct ccgtgtcagc
      961 tctgcccgcc tcagactccc ccacccccag cttcagtgag ggcctctcag acacctgtat
     1021 tcccctgcac gccagcggcc ggctgacccc ccgtgccctg cacagcttca tcaccccgcc
     1081 caccacaccc cagctgcgac ggcacaccaa gctgaagcca ccacggacgc cccccccacc
     1141 cagccgcaag gtcttccagc tgctgcccag cttccccaca ctcacccgga gcaagtccca
     1201 tgagtctcag ctggggaacc gcattgatga cgtctcctcg atgaggtttg atctctcgca
     1261 tggatcccca cagatggtac ggagggatat cgggctgtcg gtgacgcaca ggttctccac
     1321 caagtcctgg ctgtcgcagg tctgccacgt gtgccagaag agcatgatat ttggagtgaa
     1381 gtgcaagcat tgcaggttga agtgtcacaa caaatgtacc aaagaagccc ctgcctgtag
     1441 aatatccttc ctgccactaa ctcggcttcg gaggacagaa tctgtcccct cggacatcaa
     1501 caacccggtg gacagagcag ccgaacccca ttttggaacc ctccccaaag cactgacaaa
     1561 gaaggagcac cctccggcca tgaatcacct ggactccagc agcaaccctt cctccaccac
     1621 ctcctccaca ccctcctcac cggcgccctt cccgacatca tccaacccat ccagcgccac
     1681 cacgcccccc aacccctcac ctggccagcg ggacagcagg ttcaacttcc cagctgccta
     1741 cttcattcat catagacagc agtttatctt tccagtgcca tctgctggcc attgctggaa
     1801 atgcctcctt attgcagaaa gtttaaagga aaacgctttc aacatttcag cctttgcaca
     1861 cgcagccccg ctccctgaag ctgccgacgg tacccggctc gatgaccagc cgaaagcaga
     1921 tgtgttggaa gctcacgaag cggaggctga ggagccagag gctggcaagt cagaggcaga
     1981 agacgatgag gacgaggtgg acgacttgcc gagctctcgc cggccctggc ggggccccat
     2041 ctctcgcaag gccagccaga ccagcgtgta cctgcaggag tgggacatcc ccttcgagca
     2101 ggtagagctg ggcgagccca tcgggcaggg ccgctggggc cgggtgcacc gcggccgctg
     2161 gcatggcgag gtggccattc gcctgctgga gatggacggc cacaaccagg accacctgaa
     2221 gctcttcaag aaagaggtga tgaactaccg gcagacgcgg catgagaacg tggtgctctt
     2281 catgggggcc tgcatgaacc cgccccacct ggccattatc accagcttct gcaaggggcg
     2341 gacgttgcac tcgtttgtga gggaccccaa gacgtctctg gacatcaaca agacgaggca
     2401 aatcgctcag gagatcatca agggcatggg atatcttcat gccaagggca tcgtacacaa
     2461 agatctcaaa tctaagaacg tcttctatga caacggcaag gtggtcatca cagacttcgg
     2521 gctgtttggg atctcaggcg tggtccgaga gggacggcgt gagaaccagc taaagctgtc
     2581 ccacgactgg ctgtgctatc tggcccctga gattgtacgc gagatgaccc ccgggaagga
     2641 cgaggatcag ctgccattct ccaaagctgc tgatgtctat gcatttggga ctgtttggta
     2701 tgagctgcaa gcaagagact ggcccttgaa gaaccaggct gcagaggcat ccatctggca
     2761 gattggaagc ggggaaggaa tgaagcgtgt cctgacttct gtcagcttgg ggaaggaagt
     2821 cagtgagatc ctgtcggcct gctgggcttt cgacctgcag gagagaccca gcttcagcct
     2881 gctgatggac atgctggaga aacttcccaa gctgaaccgg cggctctccc accctggaca
     2941 cttctggaag tcagctgagt tgctcgagga ctacaaagac catgacggtg attataaaga
     3001 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     3061 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3121 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3181 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3241 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3301 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3361 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3421 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3481 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3541 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3601 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     3661 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3721 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3781 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3841 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3901 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     3961 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4021 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4081 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     4141 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4201 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4261 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4321 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4381 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4441 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4501 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4561 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4621 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4681 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4741 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4801 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4861 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4921 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4981 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5041 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5101 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5161 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5221 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5281 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5341 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5401 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5461 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5521 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5581 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     5641 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5701 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5761 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5821 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5881 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5941 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6001 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6061 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6121 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6181 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6241 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6781 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8101 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     8161 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     8221 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     8281 gg
//
