LOCUS       sequence-170723-        6956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for TALE-VP64 construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  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 6956)
  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..6956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1731..1740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1740..1805
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1812..1832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2615..2635
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2660..2809
                     /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             2831..2884
                     /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     3379..3399
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3563..3580)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3569..3793
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3840..3856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3840..3856)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3853..3875)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3864..3880
                     /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(3888..3894,3895..3912,3913..3918))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3933..3954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4013..4208
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4059..4194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4214..4348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4416..4433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4587..5175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4667..4686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5346..6137,6138..6206))
                     /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     5969..5988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6809..6943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttgt tgtgctgtct catcattttg gcaaagaatt cctcgaggcc gccaccatgg
     1741 actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg
     1801 acaagatggc ccccaagaag aagaggaagg tgggcattca ccgcggggta cctatggtgg
     1861 acttgaggac actcggttat tcgcaacagc aacaggagaa aatcaagcct aaggtcagga
     1921 gcaccgtcgc gcaacaccac gaggcgcttg tggggcatgg cttcactcat gcgcatattg
     1981 tcgcgctttc acagcaccct gcggcgcttg ggacggtggc tgtcaaatac caagatatga
     2041 ttgcggccct gcccgaagcc acgcacgagg caattgtagg ggtcggtaaa cagtggtcgg
     2101 gagcgcgagc acttgaggcg ctgctgactg tggcgggtga gcttaggggg cctccgctcc
     2161 agctcgacac cgggcagctg ctgaagatcg cgaagagagg gggagtaaca gcggtagagg
     2221 cagtgcacgc ctggcgcaat gcgctcaccg gggccccctt gaacctgcag gatacgttta
     2281 tcagtgataa agtgtcaagc atgacaaagt tgcagccgaa tacagtggta tcctgcaggt
     2341 gctctgtcag gaccacgggc ttacgccaca acaggtggtg gcgattgcat ctcacgacgg
     2401 cggacgccct gccttggagt ccatcgtagc ccaattgtcc aggcccgatc ccgcgttggc
     2461 tgcgttaacg aatgaccatc tggtggcgtt ggcatgtctt ggtggacgac ccgcgctcga
     2521 tgcagtcaaa aagggtctgc ctcatgctcc cgcattgatc aaaagaacca accggcggat
     2581 tcccgagaga acttcccatc gagtcgcggc tagccccaag aagaagagaa aggtggaggc
     2641 cagcggttcc ggacgggctg acgcattgga cgattttgat ctggatatgc tgggaagtga
     2701 cgccctcgat gattttgacc ttgacatgct tggttcggat gcccttgatg actttgacct
     2761 cgacatgctc ggcagtgacg cccttgatga tttcgacctg gacatgctga ttaactctag
     2821 aggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     2881 cccaaccgag tacaagccca cggtgcgcct cgccacccgc gacgaacgtc cccagggccg
     2941 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     3001 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     3061 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     3121 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     3181 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     3241 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     3301 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     3361 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     3421 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     3481 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga
     3541 tgatctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3601 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3661 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3721 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3781 gtgggctcta tggcttctga ggcggaaaga acctgcagcc caagcttggc gtaatcatgg
     3841 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3901 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3961 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     4021 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4081 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4141 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4201 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4261 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4321 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     4381 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccgcatac
     4741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     4801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5461 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5641 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5941 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6181 gcgacacgga aatgttgaat actcatactc ttcctttttc aatgaccgct ttggaaaaac
     6241 aaagactgta tttcctggaa attaatgttt attcaataaa ctgtgtattc agctatatca
     6301 catagtggtg aggctgaaat gaggcgggaa gaggcggttg gggcttaatt atatcaattt
     6361 gggtggcccc acagcgcctc caaggcgcca gtcctgtttt gacaagttgc ctctggaagc
     6421 ctctacaatg cctctcttct ttttctccag agtaagcgga ggccaggggc ccccggcctc
     6481 tgcttaatac taaaaaaaac agctgttgtc atagtaatga ttgggtggaa acattccagg
     6541 cctgggtgga gaggcttttt gcttcctctt gcaaaaccac actgacattc caggcctggg
     6601 tggagaggct ttttgcttcc tcttgcaaaa ccacactgcc ctctggaggg cagttgccta
     6661 gcaactaact aaaagaggat gtcgcacggc cagctgcggt cagttagtca cttcctgctt
     6721 aactgacttg acattttcta ttttaagagt cgggaggaaa attactgtgt tggaggccct
     6781 ccgccatctt ctgaagctga atcgaattaa cttgtttatt gcagcttata atggttacaa
     6841 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6901 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatatc atcgtc
//
