LOCUS       sequence-174791-        6666 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector for TALEN construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6666)
  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 6666)
  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: pTALEN-STAR-G
FEATURES             Location/Qualifiers
     source          1..6666
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     338..360
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     422..441
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      478..487
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             487..552
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             559..579
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1362..1949
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     misc_feature    join(2016..2590,2591..2593,2594..2602)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2183..2200)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2410..2429
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2603..3202
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2603..2622)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3099..3119
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3283..3300)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3289..3513
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3560..3576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3560..3576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3573..3595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3584..3600
                     /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(3608..3614,3615..3632,3633..3638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3653..3674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3733..3928
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3779..3914
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3934..4068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4136..4153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4307..4895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4387..4406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5066..5857,5858..5926))
                     /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     5689..5708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6519..6653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
       61 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
      121 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
      181 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
      241 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
      301 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
      361 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
      421 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcct cgaggccgcc
      481 accatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
      541 gacgatgaca agatggcccc caagaagaag aggaaggtgg gcattcaccg cggggtacct
      601 atggtggact tgaggacact cggttattcg caacagcaac aggagaaaat caagcctaag
      661 gtcaggagca ccgtcgcgca acaccacgag gcgcttgtgg ggcatggctt cactcatgcg
      721 catattgtcg cgctttcaca gcaccctgcg gcgcttggga cggtggctgt caaataccaa
      781 gatatgattg cggccctgcc cgaagccacg cacgaggcaa ttgtaggggt cggtaaacag
      841 tggtcgggag cgcgagcact tgaggcgctg ctgactgtgg cgggtgagct tagggggcct
      901 ccgctccagc tcgacaccgg gcagctgctg aagatcgcga agagaggggg agtaacagcg
      961 gtagaggcag tgcacgcctg gcgcaatgcg ctcaccgggg cccccttgaa cctgcaggat
     1021 acgtttatca gtgataaagt gtcaagcatg acaaagttgc agccgaatac agtggtatcc
     1081 tgcaggtgct ctgtcaggac cacgggctta cgccacaaca ggtggtggcg attgcatcta
     1141 atcacggcgg acgccctgcc ttggtgtcca tcgtagccca attgtccagg cccgatcccg
     1201 cgttggctgc gttaacgaat gaccatctgg tggcgttggc atgtcttggt ggacgacccg
     1261 cgctcgatgc agtcaaaaag ggtctgcctc atgctcccgc attgatcaaa agaaccaacc
     1321 ggcggattcc cgagagaact tcccatcgag tcgcgggatc ccaactagtc aaaagtgaac
     1381 tggaggagaa gaaatctgaa cttcgtcata aattgaaata tgtgcctcat gaatatattg
     1441 aattaattga aattgccaga aattccactc aggatagaat tcttgaaatg aaggtaatgg
     1501 aattttttat gaaagtttat ggatatagag gtaaacattt gggtggatca aggaaaccgg
     1561 acggagcaat ttatactgtc ggatctccta ttgattacgg tgtgatcgtg gacactaaag
     1621 cttatagcgg aggttataat ctgccaattg gccaagcaga tgaaatgcaa cgatatgtcg
     1681 aagaaaatca aacacgaaac aaacatatca accctaatga atggtggaaa gtctatccat
     1741 cttctgtaac ggaatttaag tttttatttg tgagtggtca ctttaaagga aactacaaag
     1801 ctcagcttac acgattaaat catatcacta attgtaatgg agctgttctt agtgtagaag
     1861 agcttttaat tggtggagaa atgattaaag ccggcacatt aaccttagag gaagtcagac
     1921 ggaaatttaa taacggcgag ataaacttta gatcttaaga gatcgacggt atcgagcggc
     1981 cgctcgataa gcttgatatc gaattccgcc ccccccccct ctccctcccc cccccctaac
     2041 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     2101 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     2161 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     2221 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     2281 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     2341 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     2401 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     2461 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     2521 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     2581 cgatgataat atggccacaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     2641 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     2701 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     2761 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     2821 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     2881 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     2941 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3001 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3061 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     3121 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     3181 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     3241 cgcacgaccc catgcatcga tgatctagag ctcgctgatc agcctcgact gtgccttcta
     3301 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3361 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3421 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3481 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga acctgcagcc
     3541 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3601 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3661 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3721 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3781 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3901 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     3961 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4021 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     4081 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4141 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4201 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4261 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4321 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4381 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4441 ccctgccgct taccgcatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4501 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4741 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5941 aatgaccgct ttggaaaaac aaagactgta tttcctggaa attaatgttt attcaataaa
     6001 ctgtgtattc agctatattc acatagtggt gaggctgaaa tgaggcggga agaggcggtt
     6061 ggggcttaat tatatcaatt tgggtggccc cacagcgcct ccaaggcgcc agtcctgttt
     6121 tgacaagttg cctctggaag cctctcttct ttttctccag agtaagcgga ggccaggggc
     6181 ccccggcctc tgcttaatac taaaaaaaac agctgttgtc atagtaatga ttgggtggaa
     6241 acattccagg cctgggtgga gaggcttttt gcttcctctt gcaaaaccac actgacattc
     6301 caggcctggg tggagaggct ttttgcttcc tcttgcaaaa ccacactgcc ctctggaggg
     6361 cagttgccta gcaactaatt aaaagaggat gtcgcacggc cagctgcggt cagttagtca
     6421 cttcctgctt aactgacttg acattttcta ttttaagagt cgggaggaaa attactgtgt
     6481 tggaggccct ccgccatctt ctgaagctga atcgaattaa cttgtttatt gcagcttata
     6541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6601 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatatc
     6661 atcgtc
//
