LOCUS       sequence-180160-        7236 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 7236)
  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 7236)
  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-C
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     341..363
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     425..444
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      481..490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             490..555
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             562..582
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1365..1952
                     /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(2019..2593,2594..2596,2597..2605)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2186..2203)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2413..2432
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2606..3205
                     /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(2606..2625)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3102..3122
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3286..3303)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3292..3516
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3563..3579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3563..3579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3576..3598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3587..3603
                     /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(3611..3617,3618..3635,3636..3641))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3656..3677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3736..3931
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3782..3917
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3937..4071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4310..4898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4390..4409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5069..5860,5861..5929))
                     /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     5692..5711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6522..6656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        6670..7049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
       61 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
      121 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
      181 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
      241 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
      301 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      361 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
      421 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgaggcc
      481 gccaccatgg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
      541 gatgacgatg acaagatggc ccccaagaag aagaggaagg tgggcattca ccgcggggta
      601 cctatggtgg acttgaggac actcggttat tcgcaacagc aacaggagaa aatcaagcct
      661 aaggtcagga gcaccgtcgc gcaacaccac gaggcgcttg tggggcatgg cttcactcat
      721 gcgcatattg tcgcgctttc acagcaccct gcggcgcttg ggacggtggc tgtcaaatac
      781 caagatatga ttgcggccct gcccgaagcc acgcacgagg caattgtagg ggtcggtaaa
      841 cagtggtcgg gagcgcgagc acttgaggcg ctgctgactg tggcgggtga gcttaggggg
      901 cctccgctcc agctcgacac cgggcagctg ctgaagatcg cgaagagagg gggagtaaca
      961 gcggtagagg cagtgcacgc ctggcgcaat gcgctcaccg gggccccctt gaacctgcag
     1021 gatacgttta tcagtgataa agtgtcaagc atgacaaagt tgcagccgaa tacagtggta
     1081 tcctgcaggt gctctgtcag gaccacgggc ttacgccaca acaggtggtg gcgattgcat
     1141 ctcacgacgg cggacgccct gccttggtgt ccatcgtagc ccaattgtcc aggcccgatc
     1201 ccgcgttggc tgcgttaacg aatgaccatc tggtggcgtt ggcatgtctt ggtggacgac
     1261 ccgcgctcga tgcagtcaaa aagggtctgc ctcatgctcc cgcattgatc aaaagaacca
     1321 accggcggat tcccgagaga acttcccatc gagtcgcggg atcccaacta gtcaaaagtg
     1381 aactggagga gaagaaatct gaacttcgtc ataaattgaa atatgtgcct catgaatata
     1441 ttgaattaat tgaaattgcc agaaattcca ctcaggatag aattcttgaa atgaaggtaa
     1501 tggaattttt tatgaaagtt tatggatata gaggtaaaca tttgggtgga tcaaggaaac
     1561 cggacggagc aatttatact gtcggatctc ctattgatta cggtgtgatc gtggacacta
     1621 aagcttatag cggaggttat aatctgccaa ttggccaagc agatgaaatg caacgatatg
     1681 tcgaagaaaa tcaaacacga aacaaacata tcaaccctaa tgaatggtgg aaagtctatc
     1741 catcttctgt aacggaattt aagtttttat ttgtgagtgg tcactttaaa ggaaactaca
     1801 aagctcagct tacacgatta aatcatatca ctaattgtaa tggagctgtt cttagtgtag
     1861 aagagctttt aattggtgga gaaatgatta aagccggcac attaacctta gaggaagtca
     1921 gacggaaatt taataacggc gagataaact ttagatctta agagatcgac ggtatcgagc
     1981 ggccgctcga taagcttgat atcgaattcc gccccccccc cctctccctc ccccccccct
     2041 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     2101 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     2161 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     2221 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     2281 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     2341 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     2401 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     2461 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     2521 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     2581 acacgatgat aatatggcca caaccatgac cgagtacaag cccacggtgc gcctcgccac
     2641 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     2701 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     2761 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     2821 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     2881 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     2941 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     3001 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     3061 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     3121 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     3181 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     3241 gagcgcacga ccccatgcat cgatgatcta gagctcgctg atcagcctcg actgtgcctt
     3301 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3361 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3421 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3481 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaacctgca
     3541 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3601 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3661 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3721 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3781 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3841 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3901 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     3961 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4021 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     4081 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4441 cgaccctgcc gcttaccgca tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4741 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4921 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4981 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5041 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5101 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5161 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5221 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5281 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5341 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5401 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5461 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5521 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5581 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5641 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5701 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5761 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5821 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5881 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5941 ttcaatgacc gctttggaaa aacaaagact gtatttcctg gaaattaatg tttattcaat
     6001 aaactgtgta ttcagctata ttcacatagt ggtgaggctg aaatgaggcg ggaagaggcg
     6061 gttggggctt aattatatca atttgggtgg ccccacagcg cctccaaggc gccagtcctg
     6121 ttttgacaag ttgcctctgg aagcctctct tctttttctc cagagtaagc ggaggccagg
     6181 ggcccccggc ctctgcttaa tactaaaaaa aacagctgtt gtcatagtaa tgattgggtg
     6241 gaaacattcc aggcctgggt ggagaggctt tttgcttcct cttgcaaaac cacactgaca
     6301 ttccaggcct gggtggagag gctttttgct tcctcttgca aaaccacact gccctctgga
     6361 gggcagttgc ctagcaacta attaaaagag gatgtcgcac ggccagctgc ggtcagttag
     6421 tcacttcctg cttaactgac ttgacatttt ctattttaag agtcgggagg aaaattactg
     6481 tgttggaggc cctccgccat cttctgaagc tgaatcgaat taacttgttt attgcagctt
     6541 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6601 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat
     6661 atcatcgtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     6721 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     6781 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     6841 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     6901 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     6961 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     7021 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     7081 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     7141 gtgcagcgat gggggcgggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag
     7201 gggcggggcg gggcgaggcg gagaggtgcg gcggca
//
