LOCUS       sequence-383000-        3499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU2 backbone (for TU #2 for plasmid with 2 TUs) with X2 homology 
            arms for Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3499)
  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 3499)
  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_64_TU2of2_X2homology
FEATURES             Location/Qualifiers
     source          1..3499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             356..367
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             374..1087
                     /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      1099..1170
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1186..1213
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1634..2425,2426..2494))
                     /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     2257..2276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2495..2599)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2702..3290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3191..3210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgccctgaat tcgcatctag aacggggtca tcacggctca tcatgcgcca aacaaatgtg
       61 tgcaatacac gctcggatga ctgcatgatg accgcactga ctggggacag cagatccacc
      121 taagcctgtg agagaagcag acacccgaca gatcaaggca gttacgtctc accaaaccag
      181 atgtcaacac agctacaacg tgagaccgaa agtgaaacgt gatttcatgc gtcattttga
      241 acattttgta aatcttattt aataatgtgt gcggcaattc acatttaatt tatgaatgtt
      301 ttcttaacat cgcggcaact caagaaacgg caggttcgga tcttagctac tagagaaaga
      361 ggagaaatac tagatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt
      421 ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga
      481 cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggttcc
      541 ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga
      601 ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg
      661 cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg
      721 cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagagg acggcaatat
      781 cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa
      841 acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt
      901 gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc
      961 agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga
     1021 tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact
     1081 gtacaaatga ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1141 tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt
     1201 gggcctttct gcgtttatag gtctcagctg agcatgagac ggaaatctgc tcgtcagtgg
     1261 tgctcacact gacgaatcat gtacagatca taccgatgac tgcctggcga ctcacaacta
     1321 agcaagacag ccggaaccag cgccggcgaa caccactgca tatatggcat atcacaacag
     1381 tccaactagt gcactgcagt acataacccc taggtcttcc tgagagtgac gaaggctaag
     1441 gtcacttctt cgtttccttt attggggttt ccgtgtagcc ttcccctgaa tagtgtggga
     1501 cgttttatga gaagccgtaa gaaatagccg agcggccgcg attatcaaaa aggatcttca
     1561 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1621 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1681 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1741 taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg gctccagatt
     1801 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1861 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1921 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1981 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2041 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2101 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2161 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2221 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2281 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2341 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2401 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2461 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2521 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2581 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgtcatg accaaaatcc
     2641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3361 cgttatcccc tgattctgtg gataaccgtg cggccgccaa tgtcaaaaga tcctctcata
     3421 ccatattaag taaattgcct ccatttcttt ttcctcgggc agagaaactc gcaggcaact
     3481 tgctctcgaa gtggtcacg
//
