LOCUS       sequence-174833-        5692 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 5692)
  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 5692)
  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-G
FEATURES             Location/Qualifiers
     source          1..5692
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     336..358
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     420..439
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      476..485
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             485..550
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             557..577
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1360..1380
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1405..1554
                     /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             1576..1629
                     /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     2124..2144
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2308..2325)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2314..2538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2585..2601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2585..2601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2598..2620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2609..2625
                     /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(2633..2639,2640..2657,2658..2663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2678..2699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2758..2953
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2804..2939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2959..3093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3161..3178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3332..3920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3412..3431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4091..4882,4883..4951))
                     /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     4714..4733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5544..5678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
       61 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
      121 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
      181 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
      241 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
      301 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
      361 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
      421 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcctcg aggccgccac
      481 catggactac aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga
      541 cgatgacaag atggccccca agaagaagag gaaggtgggc attcaccgcg gggtacctat
      601 ggtggacttg aggacactcg gttattcgca acagcaacag gagaaaatca agcctaaggt
      661 caggagcacc gtcgcgcaac accacgaggc gcttgtgggg catggcttca ctcatgcgca
      721 tattgtcgcg ctttcacagc accctgcggc gcttgggacg gtggctgtca aataccaaga
      781 tatgattgcg gccctgcccg aagccacgca cgaggcaatt gtaggggtcg gtaaacagtg
      841 gtcgggagcg cgagcacttg aggcgctgct gactgtggcg ggtgagctta gggggcctcc
      901 gctccagctc gacaccgggc agctgctgaa gatcgcgaag agagggggag taacagcggt
      961 agaggcagtg cacgcctggc gcaatgcgct caccggggcc cccttgaacc tgcaggatac
     1021 gtttatcagt gataaagtgt caagcatgac aaagttgcag ccgaatacag tggtatcctg
     1081 caggtgctct gtcaggacca cgggcttacg ccacaacagg tggtggcgat tgcatctaat
     1141 cacggcggac gccctgcctt ggtgtccatc gtagcccaat tgtccaggcc cgatcccgcg
     1201 ttggctgcgt taacgaatga ccatctggtg gcgttggcat gtcttggtgg acgacccgcg
     1261 ctcgatgcag tcaaaaaggg tctgcctcat gctcccgcat tgatcaaaag aaccaaccgg
     1321 cggattcccg agagaacttc ccatcgagtc gcggctagcc ccaagaagaa gagaaaggtg
     1381 gaggccagcg gttccggacg ggctgacgca ttggacgatt ttgatctgga tatgctggga
     1441 agtgacgccc tcgatgattt tgaccttgac atgcttggtt cggatgccct tgatgacttt
     1501 gacctcgaca tgctcggcag tgacgccctt gatgatttcg acctggacat gctgattaac
     1561 tctagaggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat
     1621 cctggcccaa ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga acgtccccag
     1681 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     1741 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     1801 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     1861 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     1921 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     1981 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     2041 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     2101 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     2161 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     2221 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
     2281 atcgatgatc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2341 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2401 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2461 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2521 atgcggtggg ctctatggct tctgaggcgg aaagaacctg cagcccaagc ttggcgtaat
     2581 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2641 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca
     2761 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2821 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2881 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2941 aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3001 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3061 aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg
     3121 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3181 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3241 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3481 catacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3961 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4021 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4081 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4141 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4201 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4261 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4321 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4381 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4441 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4501 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4561 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4621 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4681 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4741 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4801 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4861 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4921 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatga ccgctttgga
     4981 aaaacaaaga ctgtatttcc tggaaattaa tgtttattca ataaactgtg tattcagcta
     5041 tattcacata gtggtgaggc tgaaatgagg cgggaagagg cggttggggc ttaattatat
     5101 caatttgggt ggccccacag cgcctccaag gcgccagtcc tgttttgaca agttgcctct
     5161 ggaagcctct cttctttttc tccagagtaa gcggaggcca ggggcccccg gcctctgctt
     5221 aatactaaaa aaaacagctg ttgtcatagt aatgattggg tggaaacatt ccaggcctgg
     5281 gtggagaggc tttttgcttc ctcttgcaaa accacactga cattccaggc ctgggtggag
     5341 aggctttttg cttcctcttg caaaaccaca ctgccctctg gagggcagtt gcctagcaac
     5401 taattaaaag aggatgtcgc acggccagct gcggtcagtt agtcacttcc tgcttaactg
     5461 acttgacatt ttctatttta agagtcggga ggaaaattac tgtgttggag gccctccgcc
     5521 atcttctgaa gctgaatcga attaacttgt ttattgcagc ttataatggt tacaaataaa
     5581 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5641 tgtccaaact catcaatgta tcttatcatg tctggatctg atatcatcgt cg
//
