LOCUS       sequence-174800-        5036 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 5036)
  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 5036)
  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-C
FEATURES             Location/Qualifiers
     source          1..5036
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     337..359
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     421..440
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      477..486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             486..551
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             558..578
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1361..1381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1436..1630
                     /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(1653..1670)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1659..1883
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1930..1946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1930..1946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1943..1965)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1954..1970
                     /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(1978..1984,1985..2002,2003..2008))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2023..2044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2103..2298
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2149..2284
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2304..2438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2506..2523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2677..3265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2757..2776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3436..4227,4228..4296))
                     /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     4059..4078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4889..5023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
       61 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
      121 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
      181 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
      241 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
      301 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
      361 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
      421 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcctc gaggccgcca
      481 ccatggacta caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg
      541 acgatgacaa gatggccccc aagaagaaga ggaaggtggg cattcaccgc ggggtaccta
      601 tggtggactt gaggacactc ggttattcgc aacagcaaca ggagaaaatc aagcctaagg
      661 tcaggagcac cgtcgcgcaa caccacgagg cgcttgtggg gcatggcttc actcatgcgc
      721 atattgtcgc gctttcacag caccctgcgg cgcttgggac ggtggctgtc aaataccaag
      781 atatgattgc ggccctgccc gaagccacgc acgaggcaat tgtaggggtc ggtaaacagt
      841 ggtcgggagc gcgagcactt gaggcgctgc tgactgtggc gggtgagctt agggggcctc
      901 cgctccagct cgacaccggg cagctgctga agatcgcgaa gagaggggga gtaacagcgg
      961 tagaggcagt gcacgcctgg cgcaatgcgc tcaccggggc ccccttgaac ctgcaggata
     1021 cgtttatcag tgataaagtg tcaagcatga caaagttgca gccgaataca gtggtatcct
     1081 gcaggtgctc tgtcaggacc acgggcttac gccacaacag gtggtggcga ttgcatctca
     1141 cgacggcgga cgccctgcct tggtgtccat cgtagcccaa ttgtccaggc ccgatcccgc
     1201 gttggctgcg ttaacgaatg accatctggt ggcgttggca tgtcttggtg gacgacccgc
     1261 gctcgatgca gtcaaaaagg gtctgcctca tgctcccgca ttgatcaaaa gaaccaaccg
     1321 gcggattccc gagagaactt cccatcgagt cgcggctagc cccaagaaga agagaaaggt
     1381 ggaggccagc ggttccggac agtggatgga cgcgaaatca cttacggcat ggtcgagaac
     1441 actggttacg ttcaaggacg tgtttgtgga ctttacacgt gaggagtgga aattgctgga
     1501 tactgcgcaa caaattgtgt atcgaaatgt catgcttgag aattacaaga acctcgtcag
     1561 tctcggatac cagttgacga aaccggatgt gatccttagg ctcgaaaagg gggaagaacc
     1621 ttggctggta taaagctgag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     1681 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1741 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1801 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1861 tggggatgcg gtgggctcta tggcttctga ggcggaaaga acctgcagcc caagcttggc
     1921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     2101 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2161 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2221 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2281 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     2341 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2401 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     2461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2821 taccgcatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatgaccgct
     4321 ttggaaaaac aaagactgta tttcctggaa attaatgttt attcaataaa ctgtgtattc
     4381 agctatattc acatagtggt gaggctgaaa tgaggcggga agaggcggtt ggggcttaat
     4441 tatatcaatt tgggtggccc cacagcgcct ccaaggcgcc agtcctgttt tgacaagttg
     4501 cctctggaag cctctcttct ttttctccag agtaagcgga ggccaggggc ccccggcctc
     4561 tgcttaatac taaaaaaaac agctgttgtc atagtaatga ttgggtggaa acattccagg
     4621 cctgggtgga gaggcttttt gcttcctctt gcaaaaccac actgacattc caggcctggg
     4681 tggagaggct ttttgcttcc tcttgcaaaa ccacactgcc ctctggaggg cagttgccta
     4741 gcaactaatt aaaagaggat gtcgcacggc cagctgcggt cagttagtca cttcctgctt
     4801 aactgacttg acattttcta ttttaagagt cgggaggaaa attactgtgt tggaggccct
     4861 ccgccatctt ctgaagctga atcgaattaa cttgtttatt gcagcttata atggttacaa
     4921 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4981 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatatc atcgtc
//
