LOCUS       sequence-232410-        5068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Repair template for the TLR reporter with 10 UAS binding sites for 
            yeast Gal4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5068)
  AUTHORS   Bashir S, Dang T, Rossius J, Wolf J, Kuhn R
  TITLE     Enhancement of CRISPR-Cas9 induced precise gene editing by targeting
            histone H2A-K15 ubiquitination.
  JOURNAL   BMC Biotechnol. 2020 Oct 23;20(1):57. doi: 
            10.1186/s12896-020-00650-x.
  PUBMED    33097066
REFERENCE   2  (bases 1 to 5068)
  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: "BMC 
            Biotechnol."; date: "2020-10-23"; volume: "20(1):57. doi"; pages: " 
            10.1186/s12896-020-00650-x"
COMMENT     Sequence Label: pTLR-donor-UAS
FEATURES             Location/Qualifiers
     source          1..5068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..95
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    148..242
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     CDS             join(302..304,305..1018)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(344..365)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(605..624)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     952..973
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1027..1083
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1084..1797
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(1823..1839)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1853..1870)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1859..2083
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2177..2196)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2178..2196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2203..2220)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2203..2219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2212..2234)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2360..2815)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2497..2518)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2709..2728
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2842..2946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2947..3015,3016..3807)
                     /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     complement(3165..3184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3978..4566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4467..4486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4720..4737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4854..4875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4890..4895,4896..4913,4914..4920)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4928..4944
                     /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)."
     primer_bind     4933..4955
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4952..4968
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4952..4968
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4989..5007
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg
       61 agtactgtcc tccgagcgga gtactgtcct ccgagcggag actctagcgc tagcgcatgt
      121 cgaccgcaag cttgcatgcc tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct
      181 ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg
      241 agcggagact ctagcgctag cgcatgtcga cggtatcgat aagcttgatt aagccgccac
      301 cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      361 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      421 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      481 cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
      541 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      601 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      661 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      721 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
      781 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
      841 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
      901 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
      961 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagac
     1021 gcgttggcca cgaacttctc tctgttaaag caagcaggag atgttgaaga aaaccccggg
     1081 cctatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg
     1141 gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac
     1201 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     1261 gacatcctgg ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc
     1321 atccccgact tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca
     1381 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     1441 atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag
     1501 aaaacactcg gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa
     1561 ggcagaagcg acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag
     1621 accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg
     1681 gaccacagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag
     1741 gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacaaact taattgagcg
     1801 atcgcacgcg taaatgattg cagatccact agttctagag ctcgctgatc agcctcgact
     1861 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1921 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1981 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2041 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2101 accagctggg gctcgagatc cactagttct agcctcgagg ctagagcggc cgccaccgcg
     2161 gtggagctcc aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
     2221 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2281 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2341 cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2401 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2461 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2521 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2581 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2641 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2701 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2761 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     2821 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2881 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2941 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3061 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3121 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3181 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3241 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3301 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3361 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3481 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3541 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4621 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4681 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4741 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4801 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     4861 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     4921 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     4981 aagctcgaaa ttaaccctca ctaaagggaa caaaagctgg taccgggccc cccctcgagg
     5041 tcgaccgcaa gcttgcatgc ctgcaggt
//
