LOCUS       sequence-174762-        7131 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 7131)
  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 7131)
  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-A
FEATURES             Location/Qualifiers
     source          1..7131
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     320..342
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     404..423
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      460..469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             469..534
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             541..561
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1344..1931
                     /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    2010..2562
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2154..2171)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2381..2400
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2574..3173
                     /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(2574..2593)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3070..3090
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3254..3271)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3260..3484
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3531..3547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3531..3547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3544..3566)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3555..3571
                     /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(3579..3585,3586..3603,3604..3609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3624..3645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3704..3899
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3750..3885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3905..4039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4107..4124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4278..4866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4358..4377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5037..5828,5829..5897))
                     /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     5660..5679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6490..6624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        6638..7017
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
       61 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
      121 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
      181 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
      241 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
      301 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
      361 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
      421 gtgctgtctc atcattttgg caaagaattc ctcgaggccg ccaccatgga ctacaaagac
      481 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagatggcc
      541 cccaagaaga agaggaaggt gggcattcac cgcggggtac ctatggtgga cttgaggaca
      601 ctcggttatt cgcaacagca acaggagaaa atcaagccta aggtcaggag caccgtcgcg
      661 caacaccacg aggcgcttgt ggggcatggc ttcactcatg cgcatattgt cgcgctttca
      721 cagcaccctg cggcgcttgg gacggtggct gtcaaatacc aagatatgat tgcggccctg
      781 cccgaagcca cgcacgaggc aattgtaggg gtcggtaaac agtggtcggg agcgcgagca
      841 cttgaggcgc tgctgactgt ggcgggtgag cttagggggc ctccgctcca gctcgacacc
      901 gggcagctgc tgaagatcgc gaagagaggg ggagtaacag cggtagaggc agtgcacgcc
      961 tggcgcaatg cgctcaccgg ggcccccttg aacctgcagg atacgtttat cagtgataaa
     1021 gtgtcaagca tgacaaagtt gcagccgaat acagtggtat cctgcaggtg ctctgtcagg
     1081 accacgggct tacgccacaa caggtggtgg cgattgcatc taacattggc ggacgccctg
     1141 ccttggtgtc catcgtagcc caattgtcca ggcccgatcc cgcgttggct gcgttaacga
     1201 atgaccatct ggtggcgttg gcatgtcttg gtggacgacc cgcgctcgat gcagtcaaaa
     1261 agggtctgcc tcatgctccc gcattgatca aaagaaccaa ccggcggatt cccgagagaa
     1321 cttcccatcg agtcgcggga tcccaactag tcaaaagtga actggaggag aagaaatctg
     1381 aacttcgtca taaattgaaa tatgtgcctc atgaatatat tgaattaatt gaaattgcca
     1441 gaaattccac tcaggataga attcttgaaa tgaaggtaat ggaatttttt atgaaagttt
     1501 atggatatag aggtaaacat ttgggtggat caaggaaacc ggacggagca atttatactg
     1561 tcggatctcc tattgattac ggtgtgatcg tggacactaa agcttatagc ggaggttata
     1621 atctgccaat tggccaagca gatgaaatgc aacgatatgt cgaagaaaat caaacacgaa
     1681 acaaacatat caaccctaat gaatggtgga aagtctatcc atcttctgta acggaattta
     1741 agtttttatt tgtgagtggt cactttaaag gaaactacaa agctcagctt acacgattaa
     1801 atcatatcac taattgtaat ggagctgttc ttagtgtaga agagctttta attggtggag
     1861 aaatgattaa agccggcaca ttaaccttag aggaagtcag acggaaattt aataacggcg
     1921 agataaactt tagatcttaa gagatcgacg gtatcgagcg gccgctcgat aagcttgata
     1981 tcgaattccg cccccccccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     2041 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     2101 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     2161 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     2221 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     2281 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     2341 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     2401 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     2461 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     2521 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatgaccg
     2581 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     2641 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     2701 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     2761 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     2821 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     2881 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     2941 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     3001 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     3061 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     3121 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     3181 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg atgatctaga
     3241 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3301 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3361 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3421 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3481 atggcttctg aggcggaaag aacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     3541 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3601 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3661 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
     3721 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3901 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3961 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4021 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4081 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4141 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4201 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4261 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4321 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4381 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccgcata cctgtccgcc
     4441 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4501 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4561 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4621 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4681 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4741 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4801 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4861 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4921 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4981 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5041 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5101 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5161 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5221 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5281 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5341 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5401 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5461 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5521 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5581 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5641 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5701 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5761 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5821 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5881 aaatgttgaa tactcatact cttccttttt caatgaccgc tttggaaaaa caaagactgt
     5941 atttcctgga aattaatgtt tattcaataa actgtgtatt cagctatatt cacatagtgg
     6001 tgaggctgaa atgaggcggg aagaggcggt tggggcttaa ttatatcaat ttgggtggcc
     6061 ccacagcgcc tccaaggcgc cagtcctgtt ttgacaagtt gcctctggaa gcctctcttc
     6121 tttttctcca gagtaagcgg aggccagggg cccccggcct ctgcttaata ctaaaaaaaa
     6181 cagctgttgt catagtaatg attgggtgga aacattccag gcctgggtgg agaggctttt
     6241 tgcttcctct tgcaaaacca cactgacatt ccaggcctgg gtggagaggc tttttgcttc
     6301 ctcttgcaaa accacactgc cctctggagg gcagttgcct agcaactaat taaaagagga
     6361 tgtcgcacgg ccagctgcgg tcagttagtc acttcctgct taactgactt gacattttct
     6421 attttaagag tcgggaggaa aattactgtg ttggaggccc tccgccatct tctgaagctg
     6481 aatcgaatta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6541 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6601 aatgtatctt atcatgtctg gatctgatat catcgtcgac attgattatt gactagttat
     6661 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6721 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6781 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6841 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6901 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6961 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     7021 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     7081 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg g
//
