LOCUS       sequence-274582-        7452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express tTALE-EmGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7452)
  AUTHORS   Shin E, Kim W, Lee S, Bae J, Kim S, Ko W, Seo HS, Lim S, Lee HS, Jo 
            K
  TITLE     Truncated TALE-FP as DNA Staining Dye in a High-salt Buffer.
  JOURNAL   Sci Rep. 2019 Nov 20;9(1):17197. doi: 10.1038/s41598-019-53722-0.
  PUBMED    31748571
REFERENCE   2  (bases 1 to 7452)
  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: "2019-11-20"; volume: "9(1):17197. doi"; pages: " 
            10.1038/s41598-019-53722-0"
COMMENT     Sequence Label: pET-tTALE-EmGFP-15b
FEATURES             Location/Qualifiers
     source          1..7452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(1183..1185,1186..1188,1189..1902)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1228..1249)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1489..1508)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1836..1857
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1953..1971)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1967..2014
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2189..2194,2195..2211,2212..2217))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2244..2262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2330..2434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2435..2503,2504..3295)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2653..2672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3466..4054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3955..3974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4208..4225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4240..4379
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4305..4324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4465..4487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4481..4672)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5984..7066)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7028..7047
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7067..7144)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7350..7369
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg gatctacgca cgctcggcta cagccagcag
      181 caacaggaga agatcaaacc gaaggttcgt tcgacagtgg cgcagcacca cgaggcactg
      241 gtcggccatg ggtttacaca cgcgcacatc gttgcgctca gccaacaccc ggcagcgtta
      301 gggaccgtcg ctgtcaagta tcaggacatg atcgcagcgt tgccagaggc gacacacgaa
      361 gcgatcgttg gcgtcggcaa acagtggtcc ggcgcacgcg ctctggaggc cttgctcacg
      421 gtggcgggag agttgagagg tccaccgtta cagttggaca caggccaact tctcaagatt
      481 gcaaaacgtg gcggcgtgac cgcagtggag gcagtgcatg catggcgcaa tgcactgacg
      541 ggtgcccccc tgaacctgac tccagcacag gtggtggcta ttgcctccaa taatggggga
      601 aaacaggctc tcgaaacagt tcaaagactt ctgcctgtct tgtgccaaga tcatggactt
      661 acacccgcac aagtcgttgc aatcgctagc aatggcggtg ggaagcaagc cctggagaca
      721 gttcaaaggc tccttccagt cctgtgtcag gcacacgggc ttacccctga tcaagtggtc
      781 gcaattgcct cacatgacgg cgggaaacag gcattggaaa ctgtgcagcg cctgctcccc
      841 gttctttgcc aggatcatgg gttgacacct gcacaagttg ttgcaattgc aagcaacggc
      901 ggcggaaaac aggcactgga aactgtgcaa agactcctgc cagttttgtg ccaggctcat
      961 ggcctgactc cagaccaggt cgttgctatt gccagcaata acggaggtaa gcaagcattg
     1021 gagactgttc agcggctcct ccctgtcctc tgtcaggccc acgggctgac tcctgcacag
     1081 gtggtggcca tcgcttcaaa cggggggggg aagcaagccc tcgaaactgt tcaacggttg
     1141 ttgcctgttt tgtgtcagga tcatgggggc ggctctggcg gcatggtgag caagggcgag
     1201 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1261 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1321 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcgcc
     1381 tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1441 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct ccttcaagga cgacggcaac
     1501 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1561 aagggcaccg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1621 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1681 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1741 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1801 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1861 gccgccggga tcactctcgg catggacgag ctgtacaagt aaggatccgg ctgctaacaa
     1921 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1981 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     2041 tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg
     2101 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
     2161 tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca
     2221 tgagaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2281 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2341 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2401 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2461 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2521 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2581 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2641 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     2701 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2761 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2821 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2881 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2941 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     3001 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3061 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3121 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3181 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3241 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3301 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3361 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3421 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3481 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3541 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3601 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3661 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3721 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3781 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3841 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3901 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3961 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4021 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4081 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4141 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4201 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4261 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     4321 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     4381 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4441 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4501 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4561 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     4621 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     4681 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     4741 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     4801 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     4861 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     4921 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     4981 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     5041 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     5101 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg
     5161 agatgcgccg cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt
     5221 tggtttgcgc attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga
     5281 atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc
     5341 gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc
     5401 gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga
     5461 agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac
     5521 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
     5581 cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc
     5641 gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     5701 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     5761 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     5821 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     5881 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     5941 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     6001 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6061 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     6121 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     6181 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     6241 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     6301 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     6361 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     6421 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     6481 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     6541 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     6601 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     6661 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     6721 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     6781 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     6841 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     6901 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     6961 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     7021 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     7081 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     7141 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     7201 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     7261 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     7321 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     7381 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     7441 atcccgcgaa at
//
