LOCUS       sequence-345378-        6753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the firefly luciferase reporter upon binding of a 
            transcriptional activator to target sites. The plasmid contains 10 
            copies of target sites for the TALEN1257 DNA-binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6753)
  AUTHORS   Mesko M, Lebar T, Dekleva P, Jerala R, Bencina M
  TITLE     Engineering and Rewiring of a Calcium-Dependent Signaling Pathway.
  JOURNAL   ACS Synth Biol. 2020 Aug 21;9(8):2055-2065. doi: 
            10.1021/acssynbio.0c00133. Epub 2020 Jul 20.
  PUBMED    32643923
REFERENCE   2  (bases 1 to 6753)
  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"; doi: 
            "10.1021/acssynbio.0c00133"; journalName: "ACS Synth Biol"; date: 
            "2020-08-21- 21"; volume: "9"; issue: "8"; pages: "2055-2065"
COMMENT     Sequence Label: 10TALE_Pmin_fLuc
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    122..170
                     /note="synthetic polyadenylation signal"
     misc_feature    184..275
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     224..243
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1041..1072
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1102..1111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1108..2757
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2764..2811
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             2815..2934
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2983..3104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2994..3013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3048..3067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3298..3655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3298..3489
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3321..3341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3506..3641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3568..3587
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3686..4723
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4702..4710
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4747..4795
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4951..4968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5122..5710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5202..5221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5910..6701,6702..6753,1..17))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 aaatgttgga tgctcatact cgtccttttt caatattatt gaagcattta tcagggttac
       61 tagtacgtct ctcaaggata agtaagtaat attaaggtac gggaggtatt ggacaggccg
      121 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
      181 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
      241 ccccagtgca agtgcaggtg ccagaacatt tctctctgct tagggttagg cgttttgcgc
      301 tgctgaattc ggtggcaccg gtttgattgc acagctttac tgctgctccc gcttcttccg
      361 tttccacatc ttgctggcag ccggggtacc gcaggcaagt cgctttactg ctgctcccgc
      421 ttcttccgtt tccacatcta tggaatcacg gcggccgccc gtatctcatc ctttactgct
      481 gctcccgctt cttccgtttc cacatctaat gcgtcggcgt gcggccgctg ccagttgcat
      541 ttactgctgc tcccgcttct tccgtttcca catctttggc tgtcctaggg tacctggacg
      601 ctccttgttt actgctgctc ccgcttcttc cgtttccaca tcttggctgg tgccgcaccg
      661 gtaagcagaa ctagaggtgg caccggtttg attgcacagc tttactgctg ctcccgcttc
      721 ttccgtttcc acatcttgct ggcagccggg gtaccgcagg caagtcgctt tactgctgct
      781 cccgcttctt ccgtttccac atctatggaa tcacggcggc cgcccgtatc tcatccttta
      841 ctgctgctcc cgcttcttcc gtttccacat ctaatgcgtc ggcgtgcggc cgctgccagt
      901 tgcatttact gctgctcccg cttcttccgt ttccacatct ttggctgtcc tagggtacct
      961 ggacgctcct tgtttactgc tgctcccgct tcttccgttt ccacatcttg gctggtgccg
     1021 caccggtaag cagaggatct tagagggtat ataatggaag ctcgacttcc agctcgaggg
     1081 caatccggta ctgttggtaa agccaccatg gaagatgcca aaaacattaa gaagggccca
     1141 gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag
     1201 cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt
     1261 acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg
     1321 ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc
     1381 gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag
     1441 cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa
     1501 gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc
     1561 atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat
     1621 ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc
     1681 atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg
     1741 caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc
     1801 atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc
     1861 acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag
     1921 ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta
     1981 tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag
     2041 atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc
     2101 cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc
     2161 acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct
     2221 aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc
     2281 gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc
     2341 atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac
     2401 ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca
     2461 gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc
     2521 ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa
     2581 accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag
     2641 ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac
     2701 gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgaat
     2761 tctgcttgca agaactggtt cagtagctta agccactttg tgatccacct taacagccac
     2821 ggcttccctc ccgaggtgga ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag
     2881 gagagcggca tggatagaca ccctgctgct tgcgccagcg ccaggatcaa cgtctaactg
     2941 cagtctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     3001 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3061 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3121 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     3181 tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc ttccgctgat
     3241 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag ctaccttctg
     3301 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3361 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3421 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3481 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3541 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3601 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3661 cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga
     3721 aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga
     3781 agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag
     3841 ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact
     3901 acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag
     3961 tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt
     4021 acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg
     4081 cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg
     4141 cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc
     4201 cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg
     4261 ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg gccgcatcac
     4321 cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg tggccaacat
     4381 cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact tcgagcgccg
     4441 gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca tcggcctgga
     4501 tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg ctcaaggccg
     4561 ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg
     4621 gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg
     4681 gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag aaccggtcat
     4741 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgttcga
     4801 actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     4861 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     4921 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4981 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5041 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5101 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5161 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5221 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5281 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5341 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5401 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5461 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5521 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5581 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5641 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5701 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5761 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5821 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5881 cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga ggcaccgatc
     5941 tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt gtagatcact
     6001 acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt
     6061 tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga gcgaagaagt
     6121 ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga tgctagagta
     6181 agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg catcgtggta
     6241 tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc aagccgggtc
     6301 acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc gatcgttgtc
     6361 agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca caattctctt
     6421 accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac caagtcgttt
     6481 tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg ggacaacacc
     6541 gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc ggggcggaaa
     6601 gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct tgcacccagt
     6661 tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa
     6721 aatgccgcaa agaagggaat gagtgcgaca cga
//
