LOCUS       sequence-174826-        5818 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 5818)
  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 5818)
  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-A
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     465..487
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     549..568
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      605..614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             686..706
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1489..1509
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1534..1683
                     /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             1705..1758
                     /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     2253..2273
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2437..2454)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2443..2667
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2714..2730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2714..2730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2727..2749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2738..2754
                     /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(2762..2768,2769..2786,2787..2792))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2807..2828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2887..3082
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2933..3068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3088..3222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3290..3307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3461..4049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3541..3560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4220..5011,5012..5080))
                     /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     4843..4862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5673..5807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
       61 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
      121 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
      181 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
      241 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
      301 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
      361 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
      421 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
      481 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
      541 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga
      601 ggccgccacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaaga tggcccccaa gaagaagagg aaggtgggca ttcaccgcgg
      721 ggtacctatg gtggacttga ggacactcgg ttattcgcaa cagcaacagg agaaaatcaa
      781 gcctaaggtc aggagcaccg tcgcgcaaca ccacgaggcg cttgtggggc atggcttcac
      841 tcatgcgcat attgtcgcgc tttcacagca ccctgcggcg cttgggacgg tggctgtcaa
      901 ataccaagat atgattgcgg ccctgcccga agccacgcac gaggcaattg taggggtcgg
      961 taaacagtgg tcgggagcgc gagcacttga ggcgctgctg actgtggcgg gtgagcttag
     1021 ggggcctccg ctccagctcg acaccgggca gctgctgaag atcgcgaaga gagggggagt
     1081 aacagcggta gaggcagtgc acgcctggcg caatgcgctc accggggccc ccttgaacct
     1141 gcaggatacg tttatcagtg ataaagtgtc aagcatgaca aagttgcagc cgaatacagt
     1201 ggtatcctgc aggtgctctg tcaggaccac gggcttacgc cacaacaggt ggtggcgatt
     1261 gcatctaaca ttggcggacg ccctgccttg gtgtccatcg tagcccaatt gtccaggccc
     1321 gatcccgcgt tggctgcgtt aacgaatgac catctggtgg cgttggcatg tcttggtgga
     1381 cgacccgcgc tcgatgcagt caaaaagggt ctgcctcatg ctcccgcatt gatcaaaaga
     1441 accaaccggc ggattcccga gagaacttcc catcgagtcg cggctagccc caagaagaag
     1501 agaaaggtgg aggccagcgg ttccggacgg gctgacgcat tggacgattt tgatctggat
     1561 atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt
     1621 gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg
     1681 ctgattaact ctagaggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc
     1741 gaggagaatc ctggcccaac cgagtacaag cccacggtgc gcctcgccac ccgcgacgaa
     1801 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     1861 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     1921 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     1981 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     2041 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     2101 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     2161 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     2221 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     2281 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     2341 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     2401 accccatgca tcgatgatct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     2461 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2521 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2581 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2641 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaacctgc agcccaagct
     2701 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2761 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2821 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2881 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2941 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3001 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3061 tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa
     3121 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3181 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg
     3241 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3301 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3361 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3421 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3481 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3541 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3601 cgcttaccgc atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3661 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3721 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3781 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3841 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3901 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3961 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4021 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4081 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4141 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4201 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4261 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4321 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4381 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4441 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4501 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4561 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4621 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4681 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4741 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4801 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4861 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4921 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4981 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5041 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatgac
     5101 cgctttggaa aaacaaagac tgtatttcct ggaaattaat gtttattcaa taaactgtgt
     5161 attcagctat attcacatag tggtgaggct gaaatgaggc gggaagaggc ggttggggct
     5221 taattatatc aatttgggtg gccccacagc gcctccaagg cgccagtcct gttttgacaa
     5281 gttgcctctg gaagcctctc ttctttttct ccagagtaag cggaggccag gggcccccgg
     5341 cctctgctta atactaaaaa aaacagctgt tgtcatagta atgattgggt ggaaacattc
     5401 caggcctggg tggagaggct ttttgcttcc tcttgcaaaa ccacactgac attccaggcc
     5461 tgggtggaga ggctttttgc ttcctcttgc aaaaccacac tgccctctgg agggcagttg
     5521 cctagcaact aattaaaaga ggatgtcgca cggccagctg cggtcagtta gtcacttcct
     5581 gcttaactga cttgacattt tctattttaa gagtcgggag gaaaattact gtgttggagg
     5641 ccctccgcca tcttctgaag ctgaatcgaa ttaacttgtt tattgcagct tataatggtt
     5701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5761 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tatcatcg
//
