LOCUS       sequence-170714-        7208 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Destination vector for domain insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  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 7208)
  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-G_RFP entry
FEATURES             Location/Qualifiers
     source          1..7208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1731..1740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1740..1805
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1812..1832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2615..2635
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2671..2692
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    2793..2814
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2829..2834,2835..2852,2853..2859)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2867..2883
                     /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             2901..2912
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2919..3596
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3641..3712
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3728..3755
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3762..3782
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(3815..3832)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3821..4045
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4092..4108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4092..4108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4116..4132
                     /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(4140..4146,4147..4164,4165..4170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4185..4206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4265..4460
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4311..4446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4466..4600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4668..4685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4839..5427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4919..4938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5598..6389,6390..6458))
                     /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     6221..6240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7061..7195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttgt tgtgctgtct catcattttg gcaaagaatt cctcgaggcc gccaccatgg
     1741 actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg
     1801 acaagatggc ccccaagaag aagaggaagg tgggcattca ccgcggggta cctatggtgg
     1861 acttgaggac actcggttat tcgcaacagc aacaggagaa aatcaagcct aaggtcagga
     1921 gcaccgtcgc gcaacaccac gaggcgcttg tggggcatgg cttcactcat gcgcatattg
     1981 tcgcgctttc acagcaccct gcggcgcttg ggacggtggc tgtcaaatac caagatatga
     2041 ttgcggccct gcccgaagcc acgcacgagg caattgtagg ggtcggtaaa cagtggtcgg
     2101 gagcgcgagc acttgaggcg ctgctgactg tggcgggtga gcttaggggg cctccgctcc
     2161 agctcgacac cgggcagctg ctgaagatcg cgaagagagg gggagtaaca gcggtagagg
     2221 cagtgcacgc ctggcgcaat gcgctcaccg gggccccctt gaacctgcag gatacgttta
     2281 tcagtgataa agtgtcaagc atgacaaagt tgcagccgaa tacagtggta tcctgcaggt
     2341 gctctgtcag gaccacgggc ttacgccaca acaggtggtg gcgattgcat ctaatcacgg
     2401 cggacgccct gccttggtgt ccatcgtagc ccaattgtcc aggcccgatc ccgcgttggc
     2461 tgcgttaacg aatgaccatc tggtggcgtt ggcatgtctt ggtggacgac ccgcgctcga
     2521 tgcagtcaaa aagggtctgc ctcatgctcc cgcattgatc aaaagaacca accggcggat
     2581 tcccgagaga acttcccatc gagtcgcggc tagccccaag aagaagagaa aggtggaggc
     2641 cagcggttcc ggacagtaga gacgccattg gaattcgcgg ccgcttctag agcaatacgc
     2701 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2761 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     2821 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2881 caatttcaca catactagag aaagaggaga aatactagat ggcttcctcc gaagacgtta
     2941 tcaaagagtt catgcgtttc aaagttcgta tggaaggttc cgttaacggt cacgagttcg
     3001 aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag
     3061 ttaccaaagg tggtccgctg ccgttcgctt gggacatcct gtccccgcag ttccagtacg
     3121 gttccaaagc ttacgttaaa cacccggctg acatcccgga ctacctgaaa ctgtccttcc
     3181 cggaaggttt caaatgggaa cgtgttatga acttcgaaga cggtggtgtt gttaccgtta
     3241 cccaggactc ctccctgcaa gacggtgagt tcatctacaa agttaaactg cgtggtacca
     3301 acttcccgtc cgacggtccg gttatgcaga aaaaaaccat gggttgggaa gcttccaccg
     3361 aacgtatgta cccggaagac ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga
     3421 aagacggtgg tcactacgac gctgaagtta aaaccaccta catggctaaa aaaccggttc
     3481 agctgccggg tgcttacaaa accgacatca aactggacat cacctcccac aacgaagact
     3541 acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt gcttaataac
     3601 gctgatagtg ctagtgtaga tcgctactag agccaggcat caaataaaac gaaaggctca
     3661 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     3721 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagtag cggccgctgc
     3781 agtcagaccg tctcaagctg agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3841 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3901 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3961 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4021 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaacctgcag cccaagcttg
     4081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     4261 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4321 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4381 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4441 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     4501 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4561 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4981 cttaccgcat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatgaccg
     6481 ctttggaaaa acaaagactg tatttcctgg aaattaatgt ttattcaata aactgtgtat
     6541 tcagctatat cacatagtgg tgaggctgaa atgaggcggg aagaggcggt tggggcttaa
     6601 ttatatcaat ttgggtggcc ccacagcgcc tccaaggcgc cagtcctgtt ttgacaagtt
     6661 gcctctggaa gcctctacaa tgcctctctt ctttttctcc agagtaagcg gaggccaggg
     6721 gcccccggcc tctgcttaat actaaaaaaa acagctgttg tcatagtaat gattgggtgg
     6781 aaacattcca ggcctgggtg gagaggcttt ttgcttcctc ttgcaaaacc acactgacat
     6841 tccaggcctg ggtggagagg ctttttgctt cctcttgcaa aaccacactg ccctctggag
     6901 ggcagttgcc tagcaactaa ctaaaagagg atgtcgcacg gccagctgcg gtcagttagt
     6961 cacttcctgc ttaactgact tgacattttc tattttaaga gtcgggagga aaattactgt
     7021 gttggaggcc ctccgccatc ttctgaagct gaatcgaatt aacttgttta ttgcagctta
     7081 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     7141 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgata
     7201 tcatcgtc
//
