LOCUS       sequence-174801-        4991 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for TALE-KRAB construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4991)
  AUTHORS   Gogolok S, Garcia-Diaz C, Pollard SM
  TITLE     STAR: a simple TAL effector assembly reaction using isothermal 
            assembly.
  JOURNAL   Sci Rep. 2016 Sep 12;6:33209. doi: 10.1038/srep33209.
  PUBMED    27615025
REFERENCE   2  (bases 1 to 4991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-09-12"; volume: "6:33209. doi"; pages: " 
            10.1038/srep33209"
COMMENT     Sequence Label: pTALE-STAR-KRAB-A
FEATURES             Location/Qualifiers
     source          1..4991
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     292..314
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     376..395
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      432..441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             441..506
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             513..533
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1316..1336
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1391..1585
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     primer_bind     complement(1608..1625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1614..1838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1885..1901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1885..1901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1898..1920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1909..1925
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1933..1939,1940..1957,1958..1963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1978..1999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2058..2253
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2104..2239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2259..2393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2461..2478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2632..3220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2712..2731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3391..4182,4183..4251))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4014..4033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4844..4978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
       61 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc
      121 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa
      181 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc
      241 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
      301 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
      361 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
      421 tcctcgaggc cgccaccatg gactacaaag accatgacgg tgattataaa gatcatgaca
      481 tcgattacaa ggatgacgat gacaagatgg cccccaagaa gaagaggaag gtgggcattc
      541 accgcggggt acctatggtg gacttgagga cactcggtta ttcgcaacag caacaggaga
      601 aaatcaagcc taaggtcagg agcaccgtcg cgcaacacca cgaggcgctt gtggggcatg
      661 gcttcactca tgcgcatatt gtcgcgcttt cacagcaccc tgcggcgctt gggacggtgg
      721 ctgtcaaata ccaagatatg attgcggccc tgcccgaagc cacgcacgag gcaattgtag
      781 gggtcggtaa acagtggtcg ggagcgcgag cacttgaggc gctgctgact gtggcgggtg
      841 agcttagggg gcctccgctc cagctcgaca ccgggcagct gctgaagatc gcgaagagag
      901 ggggagtaac agcggtagag gcagtgcacg cctggcgcaa tgcgctcacc ggggccccct
      961 tgaacctgca ggatacgttt atcagtgata aagtgtcaag catgacaaag ttgcagccga
     1021 atacagtggt atcctgcagg tgctctgtca ggaccacggg cttacgccac aacaggtggt
     1081 ggcgattgca tctaacattg gcggacgccc tgccttggtg tccatcgtag cccaattgtc
     1141 caggcccgat cccgcgttgg ctgcgttaac gaatgaccat ctggtggcgt tggcatgtct
     1201 tggtggacga cccgcgctcg atgcagtcaa aaagggtctg cctcatgctc ccgcattgat
     1261 caaaagaacc aaccggcgga ttcccgagag aacttcccat cgagtcgcgg ctagccccaa
     1321 gaagaagaga aaggtggagg ccagcggttc cggacagtgg atggacgcga aatcacttac
     1381 ggcatggtcg agaacactgg ttacgttcaa ggacgtgttt gtggacttta cacgtgagga
     1441 gtggaaattg ctggatactg cgcaacaaat tgtgtatcga aatgtcatgc ttgagaatta
     1501 caagaacctc gtcagtctcg gataccagtt gacgaaaccg gatgtgatcc ttaggctcga
     1561 aaagggggaa gaaccttggc tggtataaag ctgagctcgc tgatcagcct cgactgtgcc
     1621 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1681 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1741 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1801 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaacctg
     1861 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1921 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1981 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2041 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2101 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2161 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2221 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     2281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     2401 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2461 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2521 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2581 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2641 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2701 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2761 tccgaccctg ccgcttaccg catacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2821 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2881 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2941 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3001 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3061 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3121 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3181 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3241 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3301 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3361 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3421 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3481 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3541 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3601 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3661 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3721 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3781 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3841 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3901 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3961 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4021 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4081 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4141 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4201 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4261 ttttcaatga ccgctttgga aaaacaaaga ctgtatttcc tggaaattaa tgtttattca
     4321 ataaactgtg tattcagcta tattcacata gtggtgaggc tgaaatgagg cgggaagagg
     4381 cggttggggc ttaattatat caatttgggt ggccccacag cgcctccaag gcgccagtcc
     4441 tgttttgaca agttgcctct ggaagcctct cttctttttc tccagagtaa gcggaggcca
     4501 ggggcccccg gcctctgctt aatactaaaa aaaacagctg ttgtcatagt aatgattggg
     4561 tggaaacatt ccaggcctgg gtggagaggc tttttgcttc ctcttgcaaa accacactga
     4621 cattccaggc ctgggtggag aggctttttg cttcctcttg caaaaccaca ctgccctctg
     4681 gagggcagtt gcctagcaac taattaaaag aggatgtcgc acggccagct gcggtcagtt
     4741 agtcacttcc tgcttaactg acttgacatt ttctatttta agagtcggga ggaaaattac
     4801 tgtgttggag gccctccgcc atcttctgaa gctgaatcga attaacttgt ttattgcagc
     4861 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4921 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg
     4981 atatcatcgt c
//
