LOCUS       sequence-174831-        6205 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for TALE-VP160 construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6205)
  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 6205)
  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-STARVP160-C
FEATURES             Location/Qualifiers
     source          1..6205
                     /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             1552..1935
                     /codon_start=1
                     /product="10 tandem repeats of the minimal activation 
                     domain of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP160
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGS
                     DALDDFDLDMLGSDALDDFDLDML"
     CDS             1957..2013
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2014..2724
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2159..2177)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2427..2446
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2822..2839)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2828..3052
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3099..3115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3099..3115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3112..3134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3123..3139
                     /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(3147..3153,3154..3171,3172..3177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3192..3213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3272..3467
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3318..3453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3473..3607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3675..3692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3846..4434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3926..3945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4605..5396,5397..5465))
                     /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     5228..5247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6058..6192
                     /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 gcatctcacg acggcggacg 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 ttccggacag tggagacggc gggctcgcgc cgacgcgctg
     1561 gacgatttcg atctcgacat gctgggttct gatgccctcg atgactttga cctggatatg
     1621 ttgggaagcg acgcattgga tgactttgat ctggacatgc tcggctccga tgctctggac
     1681 gatttcgatc tcgatatgtt agggtcagac gcactggatg atttcgacct tgatatgttg
     1741 ggaagcgatg cccttgatga tttcgacctg gacatgctcg gcagcgacgc cctggacgat
     1801 ttcgatctgg acatgctggg gtccgatgcc ttggatgatt ttgacttgga tatgctgggg
     1861 agtgatgccc tggacgactt tgacctggac atgctgggct ccgatgcgct cgatgacttc
     1921 gatttggata tgttgattaa ctctagaggc agtggagcca ctaacttctc cctgttgaaa
     1981 caagcagggg atgtcgaaga gaatcccggg ccaatggtga gcaagggcga ggaggataac
     2041 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2101 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2161 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     2221 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2281 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2341 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2401 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2461 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2521 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2581 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2641 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2701 ggcatggacg agctgtacaa gtaacgcccg ccccacgacc cgcagcgccc gaccgaaagg
     2761 agcgcacgac cccatgcatc gatgatctag agctccgtct caagctgagc tcgctgatca
     2821 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2881 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2941 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3001 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3061 gcggaaagaa cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     3121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3241 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     3301 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3361 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3421 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     3481 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3541 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3601 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3661 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3961 gtgcgctctc ctgttccgac cctgccgctt accgcatacc tgtccgcctt tctcccttcg
     4021 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4261 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4501 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4561 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4621 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4681 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4741 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4801 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4861 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4921 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4981 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5041 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5101 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5161 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5221 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5281 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5341 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5401 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5461 ctcatactct tcctttttca atgaccgctt tggaaaaaca aagactgtat ttcctggaaa
     5521 ttaatgttta ttcaataaac tgtgtattca gctatattca catagtggtg aggctgaaat
     5581 gaggcgggaa gaggcggttg gggcttaatt atatcaattt gggtggcccc acagcgcctc
     5641 caaggcgcca gtcctgtttt gacaagttgc ctctggaagc ctctcttctt tttctccaga
     5701 gtaagcggag gccaggggcc cccggcctct gcttaatact aaaaaaaaca gctgttgtca
     5761 tagtaatgat tgggtggaaa cattccaggc ctgggtggag aggctttttg cttcctcttg
     5821 caaaaccaca ctgacattcc aggcctgggt ggagaggctt tttgcttcct cttgcaaaac
     5881 cacactgccc tctggagggc agttgcctag caactaatta aaagaggatg tcgcacggcc
     5941 agctgcggtc agttagtcac ttcctgctta actgacttga cattttctat tttaagagtc
     6001 gggaggaaaa ttactgtgtt ggaggccctc cgccatcttc tgaagctgaa tcgaattaac
     6061 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6121 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6181 catgtctgga tctgatatca tcgtc
//
