LOCUS       sequence-383009-        4339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU3 backbone (for TU #3 for plasmid with 4-6 TUs) with HO homology 
            arms for Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4339)
  AUTHORS   Torello Pianale L, Olsson L
  TITLE     ScEnSor Kit for Saccharomyces cerevisiae Engineering and 
            Biosensor-Driven Investigation of the Intracellular Environment.
  JOURNAL   ACS Synth Biol. 2023 Aug 8. doi: 10.1021/acssynbio.3c00124.
  PUBMED    37552581
REFERENCE   2  (bases 1 to 4339)
  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: "ACS Synth 
            Biol. 2023 Aug 8. doi: 10.1021/acssynbio.3c00124."
COMMENT     Sequence Label: LT1_81_TU3of4-6_HOhomology
FEATURES             Location/Qualifiers
     source          1..4339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             352..363
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             370..1083
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1095..1166
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1182..1209
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2050..2841,2842..2910))
                     /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     2673..2692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2911..3015)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3118..3706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3607..3626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgaattcgc atctagaata aatcgcagcc aagtgagtga atagatgacg caccacggtc
       61 agacacggcc acatcgtatc tcacaggagc aagcgcgata ggagcactca cacatagtac
      121 ggtgatccgc tgactccttt gcccaaataa gacgtgagcc cgtctcagat gcacacactg
      181 gcttaagatg acaacgtgag accgaaagtg aaacgtgatt tcatgcgtca ttttgaacat
      241 tttgtaaatc ttatttaata atgtgtgcgg caattcacat ttaatttatg aatgttttct
      301 taacatcgcg gcaactcaag aaacggcagg ttcggatctt agctactaga gaaagaggag
      361 aaatactaga tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa
      421 ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca
      481 actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc ggttccttgg
      541 ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat
      601 atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg
      661 atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat
      721 accctggtaa accgcattga gctgaaaggc attgacttta aagaggacgg caatatcctg
      781 ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa
      841 aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag
      901 ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac
      961 aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat
     1021 atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac
     1081 aaatgaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1141 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     1201 ctttctgcgt ttataggtct cagctggttc tgagacggca gacctagacc acatgaggct
     1261 gatgtaggac agccaccagt ggcagctaat caacaccgca agatgccgat gcacgctcat
     1321 atcatcgtcg tcagcctggt agccattcga catacggatc agggaactcg aacccagtac
     1381 actagtgcac tgcagtacat aacccctagg tcttcctgag agttatcgtg ttgcatctgc
     1441 ggctttaaat tgatgtatct catcgcaggc acgggcagta cagtgccctg agcgtaggga
     1501 aaaatgaaaa aaaggatgta acttttaaca taattccagc acgcagcgat tgggtataat
     1561 gaagattgtt aagttcaaca acatgtgaaa ttcttagctc cgcaacttgc ttgaattatg
     1621 agctctaaga ttcaagaagt aaataggtca tataatggaa tgatagtagt acttgataaa
     1681 ggaaccatgt gatcttacgt tgatatgaaa ataattctct cacagaagtc agtttgtatg
     1741 actaacatag agagtaattg cttttcggat aaaatcgcac ctaacaaaga ttaccttttg
     1801 taaagcctcc agaacagcta tgaatgttcg ttattctacg cactttcttt catacatctt
     1861 gatccgcctt ttttgatgta ggttttaacg cggcaaaaca tattaatttt tctgatttgg
     1921 aaaaataaag agttagttct gccccgagcg gccgcgatta tcaaaaagga tcttcaccta
     1981 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2041 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2101 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2161 atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc cagatttatc
     2221 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2281 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2341 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2401 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2461 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2521 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2581 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2641 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2701 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2761 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2821 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2881 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2941 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3001 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gtcatgacca aaatccctta
     3061 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3181 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3241 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3301 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3361 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3421 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3481 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3541 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3601 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3661 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3721 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3781 atcccctgat tctgtggata accgtgcggc cgccaattct aattgtatcg agatcacttt
     3841 tcgtgatccg ctaatcagcg acggtcacat taggtttgcc aagtcagggt atgaaccata
     3901 cgatcagttt tcgtgaacct ggtacgtata ttgtggcgtt tgtgtatatt ttcattcttt
     3961 gacaacaatc aataccaacc tcaaatagga aaagtaataa gtttggcgtt acaccccaaa
     4021 agacgccaaa cggatcgaac ttactcaata gcaattagcg agacaaaacc tacgttaaga
     4081 cctgtaaccg atttatcaaa gcactctgcg gttctttctt gggaatatta cctggacatt
     4141 ttgtgccctc aagaaacgag gctctacgag cctgttggag cccctcagac attagccgcc
     4201 acgaatcaaa ctttttacgc gattcggccc aaatcagttt ctcacagatc attcgtagag
     4261 tgaaaaagca catcgattat ttgatacccc tttgggttaa ttactgttga ggtctttgcc
     4321 gtaagttcca tcgctgtcc
//
