LOCUS       V018398                 8972 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018398
VERSION     V018398
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             665..3643
                     /label="FLT3(NM_004119)"
                     /note="FLT3(NM_004119)"
                     /gene="FLT3"
     CDS             3659..3724
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3748..4336
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4402..4450
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4652..5080
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5094..5423
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5490..6284
                     /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    6460..6581
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6630..6646)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6654..6670
                     /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(6678..6708)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6723..6744
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7032..7620)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7791..8651)
                     /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(8652..8756)
                     /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 ttaagcttgc
      661 caccatgccg gcgttggcgc gcgacggcgg ccagctgccg ctgctcgttg ttttttctgc
      721 aatgatattt gggactatta caaatcaaga tctgcctgtg atcaagtgtg ttttaatcaa
      781 tcataagaac aatgattcat cagtggggaa gtcatcatca tatcccatgg tatcagaatc
      841 cccggaagac ctcgggtgtg cgttgagacc ccagagctca gggacagtgt acgaagctgc
      901 cgctgtggaa gtggatgtat ctgcttccat cacactgcaa gtgctggtcg acgccccagg
      961 gaacatttcc tgtctctggg tctttaagca cagctccctg aattgccagc cacattttga
     1021 tttacaaaac agaggagttg tttccatggt cattttgaaa atgacagaaa cccaagctgg
     1081 agaataccta ctttttattc agagtgaagc taccaattac acaatattgt ttacagtgag
     1141 tataagaaat accctgcttt acacattaag aagaccttac tttagaaaaa tggaaaacca
     1201 ggacgccctg gtctgcatat ctgagagcgt tccagagccg atcgtggaat gggtgctttg
     1261 cgattcacag ggggaaagct gtaaagaaga aagtccagct gttgttaaaa aggaggaaaa
     1321 agtgcttcat gaattatttg ggacggacat aaggtgctgt gccagaaatg aactgggcag
     1381 ggaatgcacc aggctgttca caatagatct aaatcaaact cctcagacca cattgccaca
     1441 attatttctt aaagtagggg aacccttatg gataaggtgc aaagctgttc atgtgaacca
     1501 tggattcggg ctcacctggg aattagaaaa caaagcactc gaggagggca actactttga
     1561 gatgagtacc tattcaacaa acagaactat gatacggatt ctgtttgctt ttgtatcatc
     1621 agtggcaaga aacgacaccg gatactacac ttgttcctct tcaaagcatc ccagtcaatc
     1681 agctttggtt accatcgtag aaaagggatt tataaatgct accaattcaa gtgaagatta
     1741 tgaaattgac caatatgaag agttttgttt ttctgtcagg tttaaagcct acccacaaat
     1801 cagatgtacg tggaccttct ctcgaaaatc atttccttgt gagcaaaagg gtcttgataa
     1861 cggatacagc atatccaagt tttgcaatca taagcaccag ccaggagaat atatattcca
     1921 tgcagaaaat gatgatgccc aatttaccaa aatgttcacg ctgaatataa gaaggaaacc
     1981 tcaagtgctc gcagaagcat cggcaagtca ggcgtcctgt ttctcggatg gatacccatt
     2041 accatcttgg acctggaaga agtgttcaga caagtctccc aactgcacag aagagatcac
     2101 agaaggagtc tggaatagaa aggctaacag aaaagtgttt ggacagtggg tgtcgagcag
     2161 tactctaaac atgagtgaag ccataaaagg gttcctggtc aagtgctgtg catacaattc
     2221 ccttggcaca tcttgtgaga cgatcctttt aaactctcca ggccccttcc ctttcatcca
     2281 agacaacatc tcattctatg caacaattgg tgtttgtctc ctcttcattg tcgttttaac
     2341 cctgctaatt tgtcacaagt acaaaaagca atttaggtat gaaagccagc tacagatggt
     2401 acaggtgacc ggctcctcag ataatgagta cttctacgtt gatttcagag aatatgaata
     2461 tgatctcaaa tgggagtttc caagagaaaa tttagagttt gggaaggtac taggatcagg
     2521 tgcttttgga aaagtgatga acgcaacagc ttatggaatt agcaaaacag gagtctcaat
     2581 ccaggttgcc gtcaaaatgc tgaaagaaaa agcagacagc tctgaaagag aggcactcat
     2641 gtcagaactc aagatgatga cccagctggg aagccacgag aatattgtga acctgctggg
     2701 ggcgtgcaca ctgtcaggac caatttactt gatttttgaa tactgttgct atggtgatct
     2761 tctcaactat ctaagaagta aaagagaaaa atttcacagg acttggacag agattttcaa
     2821 ggaacacaat ttcagttttt accccacttt ccaatcacat ccaaattcca gcatgcctgg
     2881 ttcaagagaa gttcagatac acccggactc ggatcaaatc tcagggcttc atgggaattc
     2941 atttcactct gaagatgaaa ttgaatatga aaaccaaaaa aggctggaag aagaggagga
     3001 cttgaatgtg cttacatttg aagatcttct ttgctttgca tatcaagttg ccaaaggaat
     3061 ggaatttctg gaatttaagt cgtgtgttca cagagacctg gccgccagga acgtgcttgt
     3121 cacccacggg aaagtggtga agatatgtga ctttggattg gctcgagata tcatgagtga
     3181 ttccaactat gttgtcaggg gcaatgcccg tctgcctgta aaatggatgg cccccgaaag
     3241 cctgtttgaa ggcatctaca ccattaagag tgatgtctgg tcatatggaa tattactgtg
     3301 ggaaatcttc tcacttggtg tgaatcctta ccctggcatt ccggttgatg ctaacttcta
     3361 caaactgatt caaaatggat ttaaaatgga tcagccattt tatgctacag aagaaatata
     3421 cattataatg caatcctgct gggcttttga ctcaaggaaa cggccatcct tccctaattt
     3481 gacttcgttt ttaggatgtc agctggcaga tgcagaagaa gcgatgtatc agaatgtgga
     3541 tggccgtgtt tcggaatgtc ctcacaccta ccaaaacagg cgacctttca gcagagagat
     3601 ggatttgggg ctactctctc cgcaggctca ggtcgaagat tcgaagcggc cgctcgagga
     3661 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3721 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     3781 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3841 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3901 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3961 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     4021 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     4081 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4141 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4201 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4261 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4321 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     4381 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     4441 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4501 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4561 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     4621 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4681 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4741 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4801 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4861 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4921 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4981 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     5041 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5101 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5161 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5221 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5281 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5341 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5401 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5461 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5521 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5581 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5641 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5701 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5761 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5821 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5881 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5941 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6001 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6061 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6121 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6181 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6241 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6301 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6361 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6421 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6481 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6541 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6601 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6661 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6721 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6781 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6841 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6901 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6961 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7021 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7081 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7141 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7201 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7261 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7321 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7381 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7441 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7501 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7561 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7621 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7681 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7741 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7801 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7861 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7921 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7981 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8041 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8101 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8161 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8221 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8281 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8341 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8401 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8461 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8521 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8581 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8641 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8761 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     8821 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     8881 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     8941 tgaccgacaa ttgcatgaag aatctgctta gg
//
