LOCUS       sequence-174788-        6664 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 6664)
  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 6664)
  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-T
FEATURES             Location/Qualifiers
     source          1..6664
                     /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..1947
                     /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(2014..2588,2589..2591,2592..2600)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2181..2198)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2408..2427
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2601..3200
                     /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(2601..2620)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3097..3117
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3281..3298)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3287..3511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3558..3574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3558..3574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3571..3593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3582..3598
                     /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(3606..3612,3613..3630,3631..3636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3651..3672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3731..3926
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3777..3912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3932..4066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4134..4151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4305..4893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4385..4404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5064..5855,5856..5924))
                     /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     5687..5706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6517..6651
                     /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 gcgggatccc aactagtcaa aagtgaactg
     1381 gaggagaaga aatctgaact tcgtcataaa ttgaaatatg tgcctcatga atatattgaa
     1441 ttaattgaaa ttgccagaaa ttccactcag gatagaattc ttgaaatgaa ggtaatggaa
     1501 ttttttatga aagtttatgg atatagaggt aaacatttgg gtggatcaag gaaaccggac
     1561 ggagcaattt atactgtcgg atctcctatt gattacggtg tgatcgtgga cactaaagct
     1621 tatagcggag gttataatct gccaattggc caagcagatg aaatgcaacg atatgtcgaa
     1681 gaaaatcaaa cacgaaacaa acatatcaac cctaatgaat ggtggaaagt ctatccatct
     1741 tctgtaacgg aatttaagtt tttatttgtg agtggtcact ttaaaggaaa ctacaaagct
     1801 cagcttacac gattaaatca tatcactaat tgtaatggag ctgttcttag tgtagaagag
     1861 cttttaattg gtggagaaat gattaaagcc ggcacattaa ccttagagga agtcagacgg
     1921 aaatttaata acggcgagat aaactttaga tcttaagaga tcgacggtat cgagcggccg
     1981 ctcgataagc ttgatatcga attccgcccc ccccccctct ccctcccccc cccctaacgt
     2041 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     2101 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     2161 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     2221 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     2281 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     2341 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     2401 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     2461 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     2521 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     2581 atgataatat ggccacaacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     2641 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     2701 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     2761 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     2821 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     2881 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     2941 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     3001 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     3061 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     3121 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     3181 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
     3241 cacgacccca tgcatcgatg atctagagct cgctgatcag cctcgactgt gccttctagt
     3301 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3361 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3421 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3481 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac ctgcagccca
     3541 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3601 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3661 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3721 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3781 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3841 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3901 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     3961 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4021 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     4081 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4141 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4201 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4261 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4321 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4381 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4441 ctgccgctta ccgcatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4501 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4561 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4621 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4681 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4741 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4801 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4861 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4921 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4981 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5041 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5101 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5161 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5221 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5281 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5341 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5401 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5461 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5521 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5581 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5641 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5701 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5761 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5821 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5881 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5941 tgaccgcttt ggaaaaacaa agactgtatt tcctggaaat taatgtttat tcaataaact
     6001 gtgtattcag ctatattcac atagtggtga ggctgaaatg aggcgggaag aggcggttgg
     6061 ggcttaatta tatcaatttg ggtggcccca cagcgcctcc aaggcgccag tcctgttttg
     6121 acaagttgcc tctggaagcc tctcttcttt ttctccagag taagcggagg ccaggggccc
     6181 ccggcctctg cttaatacta aaaaaaacag ctgttgtcat agtaatgatt gggtggaaac
     6241 attccaggcc tgggtggaga ggctttttgc ttcctcttgc aaaaccacac tgacattcca
     6301 ggcctgggtg gagaggcttt ttgcttcctc ttgcaaaacc acactgccct ctggagggca
     6361 gttgcctagc aactaattaa aagaggatgt cgcacggcca gctgcggtca gttagtcact
     6421 tcctgcttaa ctgacttgac attttctatt ttaagagtcg ggaggaaaat tactgtgttg
     6481 gaggccctcc gccatcttct gaagctgaat cgaattaact tgtttattgc agcttataat
     6541 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6601 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatatcat
     6661 cgtc
//
