LOCUS       sequence-174828-        5549 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for TALE-VP64 construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5549)
  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 5549)
  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-STARVP64-T
FEATURES             Location/Qualifiers
     source          1..5549
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     194..216
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     278..297
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      334..343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             343..408
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             415..435
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1218..1238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1263..1412
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             1434..1487
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     1982..2002
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2166..2183)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2172..2396
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2443..2459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2443..2459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2456..2478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2467..2483
                     /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(2491..2497,2498..2515,2516..2521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2536..2557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2616..2811
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2662..2797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2817..2951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3019..3036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3190..3778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3270..3289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3949..4740,4741..4809))
                     /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     4572..4591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5402..5536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
       61 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
      121 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
      181 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
      241 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
      301 tctcatcatt ttggcaaaga attcctcgag gccgccacca tggactacaa agaccatgac
      361 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagat ggcccccaag
      421 aagaagagga aggtgggcat tcaccgcggg gtacctatgg tggacttgag gacactcggt
      481 tattcgcaac agcaacagga gaaaatcaag cctaaggtca ggagcaccgt cgcgcaacac
      541 cacgaggcgc ttgtggggca tggcttcact catgcgcata ttgtcgcgct ttcacagcac
      601 cctgcggcgc ttgggacggt ggctgtcaaa taccaagata tgattgcggc cctgcccgaa
      661 gccacgcacg aggcaattgt aggggtcggt aaacagtggt cgggagcgcg agcacttgag
      721 gcgctgctga ctgtggcggg tgagcttagg gggcctccgc tccagctcga caccgggcag
      781 ctgctgaaga tcgcgaagag agggggagta acagcggtag aggcagtgca cgcctggcgc
      841 aatgcgctca ccggggcccc cttgaacctg caggatacgt ttatcagtga taaagtgtca
      901 agcatgacaa agttgcagcc gaatacagtg gtatcctgca ggtgctctgt caggaccacg
      961 ggcttacgcc acaacaggtg gtggcgattg catctaatgg aggcggacgc cctgccttgg
     1021 agtccatcgt agcccaattg tccaggcccg atcccgcgtt ggctgcgtta acgaatgacc
     1081 atctggtggc gttggcatgt cttggtggac gacccgcgct cgatgcagtc aaaaagggtc
     1141 tgcctcatgc tcccgcattg atcaaaagaa ccaaccggcg gattcccgag agaacttccc
     1201 atcgagtcgc ggctagcccc aagaagaaga gaaaggtgga ggccagcggt tccggacggg
     1261 ctgacgcatt ggacgatttt gatctggata tgctgggaag tgacgccctc gatgattttg
     1321 accttgacat gcttggttcg gatgcccttg atgactttga cctcgacatg ctcggcagtg
     1381 acgcccttga tgatttcgac ctggacatgc tgattaactc tagaggcagt ggagagggca
     1441 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaacc gagtacaagc
     1501 ccacggtgcg cctcgccacc cgcgacgaac gtccccaggg ccgtacgcac cctcgccgcc
     1561 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     1621 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     1681 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     1741 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     1801 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     1861 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     1921 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     1981 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     2041 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     2101 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgatgatcta gagctcgctg
     2161 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2221 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2281 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2341 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2401 tgaggcggaa agaacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     2461 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2521 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2581 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
     2641 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2701 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     2761 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
     2821 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2881 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2941 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgca tacctgtccg cctttctccc
     3361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4801 aatactcata ctcttccttt ttcaatgacc gctttggaaa aacaaagact gtatttcctg
     4861 gaaattaatg tttattcaat aaactgtgta ttcagctata ttcacatagt ggtgaggctg
     4921 aaatgaggcg ggaagaggcg gttggggctt aattatatca atttgggtgg ccccacagcg
     4981 cctccaaggc gccagtcctg ttttgacaag ttgcctctgg aagcctctct tctttttctc
     5041 cagagtaagc ggaggccagg ggcccccggc ctctgcttaa tactaaaaaa aacagctgtt
     5101 gtcatagtaa tgattgggtg gaaacattcc aggcctgggt ggagaggctt tttgcttcct
     5161 cttgcaaaac cacactgaca ttccaggcct gggtggagag gctttttgct tcctcttgca
     5221 aaaccacact gccctctgga gggcagttgc ctagcaacta attaaaagag gatgtcgcac
     5281 ggccagctgc ggtcagttag tcacttcctg cttaactgac ttgacatttt ctattttaag
     5341 agtcgggagg aaaattactg tgttggaggc cctccgccat cttctgaagc tgaatcgaat
     5401 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5521 ttatcatgtc tggatctgat atcatcgtc
//
