LOCUS       sequence-174829-        5943 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Destination vector for domain insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5943)
  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 5943)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-STAR-A_RFP entry
FEATURES             Location/Qualifiers
     source          1..5943
                     /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..1380
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1416..1437
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    1538..1559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1574..1579,1580..1597,1598..1604)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1612..1628
                     /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)."
     RBS             1646..1657
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1664..2341
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2386..2457
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2473..2500
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2507..2527
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(2560..2577)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2566..2790
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2837..2853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2837..2853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2861..2877
                     /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(2885..2891,2892..2909,2910..2915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2930..2951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3010..3205
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3056..3191
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3211..3345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3413..3430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3584..4172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3664..3683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4343..5134,5135..5203))
                     /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     4966..4985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5796..5930
                     /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 tgcatctaac
     1141 attggcggac gccctgcctt ggtgtccatc gtagcccaat tgtccaggcc cgatcccgcg
     1201 ttggctgcgt taacgaatga ccatctggtg gcgttggcat gtcttggtgg acgacccgcg
     1261 ctcgatgcag tcaaaaaggg tctgcctcat gctcccgcat tgatcaaaag aaccaaccgg
     1321 cggattcccg agagaacttc ccatcgagtc gcggctagcc ccaagaagaa gagaaaggtg
     1381 gaggccagcg gttccggaca gtagagacgc cattggaatt cgcggccgct tctagagcaa
     1441 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     1501 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     1561 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     1621 gataacaatt tcacacatac tagagaaaga ggagaaatac tagatggctt cctccgaaga
     1681 cgttatcaaa gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta acggtcacga
     1741 gttcgaaatc gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact
     1801 gaaagttacc aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc cgcagttcca
     1861 gtacggttcc aaagcttacg ttaaacaccc ggctgacatc ccggactacc tgaaactgtc
     1921 cttcccggaa ggtttcaaat gggaacgtgt tatgaacttc gaagacggtg gtgttgttac
     1981 cgttacccag gactcctccc tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg
     2041 taccaacttc ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc
     2101 caccgaacgt atgtacccgg aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa
     2161 actgaaagac ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc
     2221 ggttcagctg ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga
     2281 agactacacc atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta
     2341 ataacgctga tagtgctagt gtagatcgct actagagcca ggcatcaaat aaaacgaaag
     2401 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     2461 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agtagcggcc
     2521 gctgcagtca gaccgtctca agctgagctc gctgatcagc ctcgactgtg ccttctagtt
     2581 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2641 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2701 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2761 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc tgcagcccaa
     2821 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2881 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2941 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3001 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3061 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3121 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3181 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     3241 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3301 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     3361 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3721 tgccgcttac cgcatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4021 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4261 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4321 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4381 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4441 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4501 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4561 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4621 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4681 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4741 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4801 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4861 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4921 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4981 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5041 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5101 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5161 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5221 gaccgctttg gaaaaacaaa gactgtattt cctggaaatt aatgtttatt caataaactg
     5281 tgtattcagc tatattcaca tagtggtgag gctgaaatga ggcgggaaga ggcggttggg
     5341 gcttaattat atcaatttgg gtggccccac agcgcctcca aggcgccagt cctgttttga
     5401 caagttgcct ctggaagcct ctcttctttt tctccagagt aagcggaggc caggggcccc
     5461 cggcctctgc ttaatactaa aaaaaacagc tgttgtcata gtaatgattg ggtggaaaca
     5521 ttccaggcct gggtggagag gctttttgct tcctcttgca aaaccacact gacattccag
     5581 gcctgggtgg agaggctttt tgcttcctct tgcaaaacca cactgccctc tggagggcag
     5641 ttgcctagca actaattaaa agaggatgtc gcacggccag ctgcggtcag ttagtcactt
     5701 cctgcttaac tgacttgaca ttttctattt taagagtcgg gaggaaaatt actgtgttgg
     5761 aggccctccg ccatcttctg aagctgaatc gaattaactt gtttattgca gcttataatg
     5821 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5881 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatatcatc
     5941 gtc
//
