LOCUS       sequence-174832-        6076 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 6076)
  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 6076)
  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-T
FEATURES             Location/Qualifiers
     source          1..6076
                     /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             1423..1806
                     /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             1828..1884
                     /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             1885..2595
                     /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(2030..2048)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2298..2317
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2693..2710)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2699..2923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2970..2986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2970..2986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2983..3005)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2994..3010
                     /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(3018..3024,3025..3042,3043..3048))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3063..3084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3143..3338
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3189..3324
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3344..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3546..3563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3717..4305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3797..3816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4476..5267,5268..5336))
                     /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     5099..5118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5929..6063
                     /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 ggaggcggac gccctgcctt ggagtccatc 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 gttccggaca gtggagacgg cgggctcgcg ccgacgcgct ggacgatttc
     1441 gatctcgaca tgctgggttc tgatgccctc gatgactttg acctggatat gttgggaagc
     1501 gacgcattgg atgactttga tctggacatg ctcggctccg atgctctgga cgatttcgat
     1561 ctcgatatgt tagggtcaga cgcactggat gatttcgacc ttgatatgtt gggaagcgat
     1621 gcccttgatg atttcgacct ggacatgctc ggcagcgacg ccctggacga tttcgatctg
     1681 gacatgctgg ggtccgatgc cttggatgat tttgacttgg atatgctggg gagtgatgcc
     1741 ctggacgact ttgacctgga catgctgggc tccgatgcgc tcgatgactt cgatttggat
     1801 atgttgatta actctagagg cagtggagcc actaacttct ccctgttgaa acaagcaggg
     1861 gatgtcgaag agaatcccgg gccaatggtg agcaagggcg aggaggataa catggccatc
     1921 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     1981 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     2041 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     2101 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     2161 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     2221 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     2281 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     2341 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     2401 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     2461 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     2521 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2581 gagctgtaca agtaacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     2641 ccccatgcat cgatgatcta gagctccgtc tcaagctgag ctcgctgatc agcctcgact
     2701 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2761 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2821 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2881 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2941 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3001 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3061 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3121 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     3181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3241 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3301 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     3361 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     3481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3841 cctgttccga ccctgccgct taccgcatac ctgtccgcct ttctcccttc gggaagcgtg
     3901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4381 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4441 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4501 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4561 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4621 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4681 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4741 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4801 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4861 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4921 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4981 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5041 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5101 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5161 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5221 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5281 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5341 ttcctttttc aatgaccgct ttggaaaaac aaagactgta tttcctggaa attaatgttt
     5401 attcaataaa ctgtgtattc agctatattc acatagtggt gaggctgaaa tgaggcggga
     5461 agaggcggtt ggggcttaat tatatcaatt tgggtggccc cacagcgcct ccaaggcgcc
     5521 agtcctgttt tgacaagttg cctctggaag cctctcttct ttttctccag agtaagcgga
     5581 ggccaggggc ccccggcctc tgcttaatac taaaaaaaac agctgttgtc atagtaatga
     5641 ttgggtggaa acattccagg cctgggtgga gaggcttttt gcttcctctt gcaaaaccac
     5701 actgacattc caggcctggg tggagaggct ttttgcttcc tcttgcaaaa ccacactgcc
     5761 ctctggaggg cagttgccta gcaactaatt aaaagaggat gtcgcacggc cagctgcggt
     5821 cagttagtca cttcctgctt aactgacttg acattttcta ttttaagagt cgggaggaaa
     5881 attactgtgt tggaggccct ccgccatctt ctgaagctga atcgaattaa cttgtttatt
     5941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6061 atctgatatc atcgtc
//
