LOCUS       sequence-174843-        5033 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 5033)
  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 5033)
  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-C
FEATURES             Location/Qualifiers
     source          1..5033
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     199..221
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     283..302
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      339..348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             348..413
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             420..440
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1223..1243
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1268..1417
                     /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             1439..1492
                     /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     1987..2007
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2171..2188)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2177..2401
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2448..2464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2448..2464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2461..2483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2472..2488
                     /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(2496..2502,2503..2520,2521..2526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2541..2562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2621..2816
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2667..2802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2822..2956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2859..2878)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2913..2932
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3024..3041)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3195..3783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3275..3294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3954..4745,4746..4814))
                     /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     4577..4596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tctgtgcgga gccgamwtct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
       61 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
      121 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
      181 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
      241 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
      301 tgctgtctca tcattttggc aaagaattcc tcgaggccgc caccatggac tacaaagacc
      361 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagatggccc
      421 ccaagaagaa gaggaaggtg ggcattcacc gcggggtacc tatggtggac ttgaggacac
      481 tcggttattc gcaacagcaa caggagaaaa tcaagcctaa ggtcaggagc accgtcgcgc
      541 aacaccacga ggcgcttgtg gggcatggct tcactcatgc gcatattgtc gcgctttcac
      601 agcaccctgc ggcgcttggg acggtggctg tcaaatacca agatatgatt gcggccctgc
      661 ccgaagccac gcacgaggca attgtagggg tcggtaaaca gtggtcggga gcgcgagcac
      721 ttgaggcgct gctgactgtg gcgggtgagc ttagggggcc tccgctccag ctcgacaccg
      781 ggcagctgct gaagatcgcg aagagagggg gagtaacagc ggtagaggca gtgcacgcct
      841 ggcgcaatgc gctcaccggg gcccccttga acctgcagga tacgtttatc agtgataaag
      901 tgtcaagcat gacaaagttg cagccgaata cagtggtatc ctgcaggtgc tctgtcagga
      961 ccacgggctt acgccacaac aggtggtggc gattgcatct cacgacggcg gacgccctgc
     1021 cttggtgtcc atcgtagccc aattgtccag gcccgatccc gcgttggctg cgttaacgaa
     1081 tgaccatctg gtggcgttgg catgtcttgg tggacgaccc gcgctcgatg cagtcaaaaa
     1141 gggtctgcct catgctcccg cattgatcaa aagaaccaac cggcggattc ccgagagaac
     1201 ttcccatcga gtcgcggcta gccccaagaa gaagagaaag gtggaggcca gcggttccgg
     1261 acgggctgac gcattggacg attttgatct ggatatgctg ggaagtgacg ccctcgatga
     1321 ttttgacctt gacatgcttg gttcggatgc ccttgatgac tttgacctcg acatgctcgg
     1381 cagtgacgcc cttgatgatt tcgacctgga catgctgatt aactctagag gcagtggaga
     1441 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc caaccgagta
     1501 caagcccacg gtgcgcctcg ccacccgcga cgaacgtccc cagggccgta cgcaccctcg
     1561 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     1621 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     1681 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     1741 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     1801 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     1861 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     1921 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     1981 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     2041 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc
     2101 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcgatg atctagagct
     2161 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2221 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2281 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2341 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2401 gcttctgagg cggaaagaac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     2461 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2521 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2581 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     2641 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2701 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2761 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2821 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2881 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2941 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3001 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3061 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3121 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccgcatacct gtccgccttt
     3361 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3601 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3841 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3901 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3961 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4021 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4081 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4141 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4201 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4261 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4321 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4381 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4441 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4501 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4561 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4621 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4681 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4741 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4801 tgttgaatac tcatactctt cctttttcaa tgaccgcttt ggaaaaacaa agactgtatt
     4861 tcctggaaat taatgtttat tcaataaact gtgtattcag ctatattcac atagtggtga
     4921 ggctgaaatg aggcgggaag aggcggttgg ggcttaatta tatcaatttg ggtggcccca
     4981 cagcgcctcc aaggcgccag tcctgttttg acaagttgcc tctggaagcc tct
//
