LOCUS       sequence-174798-        5034 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 5034)
  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 5034)
  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-T
FEATURES             Location/Qualifiers
     source          1..5034
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     335..357
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     419..438
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      475..484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             484..549
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             556..576
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1359..1379
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1434..1628
                     /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(1651..1668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1657..1881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1928..1944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1928..1944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1941..1963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1952..1968
                     /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(1976..1982,1983..2000,2001..2006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2021..2042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2101..2296
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2147..2282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2302..2436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2504..2521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2675..3263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2755..2774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3434..4225,4226..4294))
                     /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     4057..4076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4887..5021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
       61 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
      121 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
      181 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
      241 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
      301 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
      361 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
      421 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga ggccgccacc
      481 atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
      541 gatgacaaga tggcccccaa gaagaagagg aaggtgggca ttcaccgcgg ggtacctatg
      601 gtggacttga ggacactcgg ttattcgcaa cagcaacagg agaaaatcaa gcctaaggtc
      661 aggagcaccg tcgcgcaaca ccacgaggcg cttgtggggc atggcttcac tcatgcgcat
      721 attgtcgcgc tttcacagca ccctgcggcg cttgggacgg tggctgtcaa ataccaagat
      781 atgattgcgg ccctgcccga agccacgcac gaggcaattg taggggtcgg taaacagtgg
      841 tcgggagcgc gagcacttga ggcgctgctg actgtggcgg gtgagcttag ggggcctccg
      901 ctccagctcg acaccgggca gctgctgaag atcgcgaaga gagggggagt aacagcggta
      961 gaggcagtgc acgcctggcg caatgcgctc accggggccc ccttgaacct gcaggatacg
     1021 tttatcagtg ataaagtgtc aagcatgaca aagttgcagc cgaatacagt ggtatcctgc
     1081 aggtgctctg tcaggaccac gggcttacgc cacaacaggt ggtggcgatt gcatctaatg
     1141 gaggcggacg ccctgccttg gagtccatcg tagcccaatt gtccaggccc gatcccgcgt
     1201 tggctgcgtt aacgaatgac catctggtgg cgttggcatg tcttggtgga cgacccgcgc
     1261 tcgatgcagt caaaaagggt ctgcctcatg ctcccgcatt gatcaaaaga accaaccggc
     1321 ggattcccga gagaacttcc catcgagtcg cggctagccc caagaagaag agaaaggtgg
     1381 aggccagcgg ttccggacag tggatggacg cgaaatcact tacggcatgg tcgagaacac
     1441 tggttacgtt caaggacgtg tttgtggact ttacacgtga ggagtggaaa ttgctggata
     1501 ctgcgcaaca aattgtgtat cgaaatgtca tgcttgagaa ttacaagaac ctcgtcagtc
     1561 tcggatacca gttgacgaaa ccggatgtga tccttaggct cgaaaagggg gaagaacctt
     1621 ggctggtata aagctgagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     1681 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1741 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     1801 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     1861 gggatgcggt gggctctatg gcttctgagg cggaaagaac ctgcagccca agcttggcgt
     1921 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1981 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2041 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     2101 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2161 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2221 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2281 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     2341 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2401 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     2461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2581 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2641 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2701 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2761 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2821 ccgcatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2881 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2941 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3001 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3061 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3121 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3181 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3241 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3301 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3361 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3421 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3481 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3541 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3601 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3661 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3721 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3781 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3841 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3901 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3961 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4021 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4081 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4141 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4201 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4261 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tgaccgcttt
     4321 ggaaaaacaa agactgtatt tcctggaaat taatgtttat tcaataaact gtgtattcag
     4381 ctatattcac atagtggtga ggctgaaatg aggcgggaag aggcggttgg ggcttaatta
     4441 tatcaatttg ggtggcccca cagcgcctcc aaggcgccag tcctgttttg acaagttgcc
     4501 tctggaagcc tctcttcttt ttctccagag taagcggagg ccaggggccc ccggcctctg
     4561 cttaatacta aaaaaaacag ctgttgtcat agtaatgatt gggtggaaac attccaggcc
     4621 tgggtggaga ggctttttgc ttcctcttgc aaaaccacac tgacattcca ggcctgggtg
     4681 gagaggcttt ttgcttcctc ttgcaaaacc acactgccct ctggagggca gttgcctagc
     4741 aactaattaa aagaggatgt cgcacggcca gctgcggtca gttagtcact tcctgcttaa
     4801 ctgacttgac attttctatt ttaagagtcg ggaggaaaat tactgtgttg gaggccctcc
     4861 gccatcttct gaagctgaat cgaattaact tgtttattgc agcttataat ggttacaaat
     4921 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4981 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatatcat cgtc
//
