LOCUS       40924_6816        4570 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBlueTAL, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4570)
  AUTHORS   Takasu Y, Sajwan S, Daimon T, Osanai-Futahashi M, Uchino K, Sezutsu 
            H, Tamura T, Zurovec M.
  TITLE     Efficient TALEN construction for Bombyx mori gene targeting
  JOURNAL   PLoS ONE 8 (9), E73458 (2013)
  PUBMED    24058473
REFERENCE   2  (bases 1 to 4570)
  AUTHORS   Takasu Y, Sajwan S, Daimon T, Osanai-Futahashi M, Uchino K, Sezutsu 
            H, Tamura T, Zurovec M.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-OCT-2013) Genetics, Biology Centre, CAS, Institute of 
            Entomology, Branisovska 31, Ceske Budejovice 37005, Czech Republic
REFERENCE   3  (bases 1 to 4570)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4570)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2013"; volume: "8"; issue: "9"; pages: "E73458"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-OCT-2013) Genetics, Biology Centre, CAS, Institute of 
            Entomology, Branisovska 31, Ceske Budejovice 37005, Czech Republic"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        542..560
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             640..705
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             712..732
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     misc_feature    complement(1159..1164)
                     /label=BsmBI restriction site
                     /note="BsmBI restriction site"
     primer_bind     1326..1342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1352..1370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1415..1433)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1454..1470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1478..1494)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1502..1532)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1547..1568)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    1603..1608
                     /label=BsmBI restriction site
                     /note="BsmBI restriction site"
     CDS             1754..2341
                     /codon_start=1
                     /label=FokI cleavage domain
                     /note="nonspecific DNA cleavage domain of the FokI
                     endonuclease (Li et al., 1992)"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     polyA_signal    2401..2520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2826..3414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3588..4445)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4446..4550)
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc aggcagtcga
      541 ctaatacgac tcactatagg gcgcgcctca tttactaagg tgtgctcgaa cagtgcgcat
      601 tcgcatctac gtacttgtca cttattaaaa atcaaaatgg attacaagga ccacgacggc
      661 gactacaagg accacgacat cgactacaag gacgacgacg acaagatggc tcccaagaag
      721 aagcgtaaag tgggcatcca cggagtccct atggttgact tgcgcactct gggctactcc
      781 cagcaacagc aagagaagat caagcccaag gtgcgctcta ccgtcgctca gcaccacgaa
      841 gccctggttg gtcacggctt cactcacgct cacatcgtgg ctctgagcca gcacccagct
      901 gctctcggaa cagtggctgt caagtaccaa gatatgatcg ctgccttgcc ggaggctacc
      961 cacgaagcca tcgttggagt gggcaagcag tggtccggtg ctcgtgctct ggaggctctg
     1021 ctgactgtgg ctggtgaact cagaggacca cctttgcaac tggatacagg acagttgctg
     1081 aagatcgcca agaggggagg agtgaccgcc gttgaagccg tgcatgcttg gcgtaacgcc
     1141 ctgaccggtg ctcccctgga gacgggcgcc gctacagggc gcgtcccatt cgccattcag
     1201 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     1261 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     1321 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
     1381 taccgggccc ccccccgagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     1441 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1501 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1561 tcacattaat tgcgttgcgc tcactgcccg ctttccaccg gtcgtctcca acgaccacct
     1621 cgtggctctg gcctgcctgg gtggccgtcc cgctctggac gctgtcaaga agggactccc
     1681 ccacgctcct gccttgatca agcgcaccaa ccgtcgcatc cccgagagga cttcccacag
     1741 agtggctgga tcccagttgg ttaagagcga actggaggaa aagaagtcag agctgaggca
     1801 caagctcaag tacgtgcctc acgagtacat cgaactcatc gagatcgcta ggaactccac
     1861 ccaagacaga atcttggaaa tgaaggtcat ggagttcttc atgaaggtct acggttaccg
     1921 tggcaagcac ctcggtggct cgcgcaagcc agatggagcc atctacactg tgggttcccc
     1981 gatcgactac ggtgtgatcg tcgatacaaa ggcttactcc ggaggttaca acctgcccat
     2041 cggccaggcc gacgagatgc aacgttacgt cgaggaaaac cagaccagaa acaagcacat
     2101 caaccccaac gaatggtgga aggtctaccc ttcctctgtt actgagttca agttcctgtt
     2161 cgtttctgga cacttcaagg gtaactacaa ggctcaactc acacgcttga accacatcac
     2221 caactgcaac ggtgccgttc tcagcgtgga ggaactgctc atcggcggag aaatgattaa
     2281 ggctggcacc ttgactctgg aggaagtccg tcgcaagttc aacaacggag agatcaactt
     2341 cagatcttaa ctcgagtccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2401 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2461 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2521 cgagtttaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaat ctagatgcat tcgcgaggta
     2581 ccgagctcgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2641 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2701 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2761 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2821 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2881 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2941 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3001 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3061 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3241 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4441 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4501 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4561 cgaaaagtgc
//
