LOCUS       sequence-382891-        4339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU4 backbone (for TU #4 for plasmid with 4 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_82_TU4of4_HOhomology
FEATURES             Location/Qualifiers
     source          1..4339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             355..366
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             373..1086
                     /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      1098..1169
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1185..1212
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2053..2844,2845..2913))
                     /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     2676..2695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2914..3018)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3121..3709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3610..3629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccctgaatt cgcatctaga gcagacctag accacatgag gctgatgtag gacagccacc
       61 agtggcagct aatcaacacc gcaagatgcc gatgcacgct catatcatcg tcgtcagcct
      121 ggtagccatt cgacatacgg atcagggaac tcgaacccag taccgtctca gttctgtatg
      181 ggcacagaca acctaaacgt gagaccgaaa gtgaaacgtg atttcatgcg tcattttgaa
      241 cattttgtaa atcttattta ataatgtgtg cggcaattca catttaattt atgaatgttt
      301 tcttaacatc gcggcaactc aagaaacggc aggttcggat cttagctact agagaaagag
      361 gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg
      421 gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac
      481 gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggttcct
      541 tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac
      601 catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc
      661 acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc
      721 gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagagga cggcaatatc
      781 ctgggccata agctggaata caattttaac agccacaatg tttacatcac cgccgataaa
      841 caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg
      901 cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca
      961 gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat
     1021 catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg
     1081 tacaaatgac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1141 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     1201 ggcctttctg cgtttatagg tctcagctga gcatgagacg gaaatctgct cgtcagtggt
     1261 gctcacactg acgaatcatg tacagatcat accgatgact gcctggcgac tcacaactaa
     1321 gcaagacagc cggaaccagc gccggcgaac accactgcat atatggcata tcacaacagt
     1381 ccaactagtg cactgcagta cataacccct aggtcttcct gagagttatc gtgttgcatc
     1441 tgcggcttta aattgatgta tctcatcgca ggcacgggca gtacagtgcc ctgagcgtag
     1501 ggaaaaatga aaaaaaggat gtaactttta acataattcc agcacgcagc gattgggtat
     1561 aatgaagatt gttaagttca acaacatgtg aaattcttag ctccgcaact tgcttgaatt
     1621 atgagctcta agattcaaga agtaaatagg tcatataatg gaatgatagt agtacttgat
     1681 aaaggaacca tgtgatctta cgttgatatg aaaataattc tctcacagaa gtcagtttgt
     1741 atgactaaca tagagagtaa ttgcttttcg gataaaatcg cacctaacaa agattacctt
     1801 ttgtaaagcc tccagaacag ctatgaatgt tcgttattct acgcactttc tttcatacat
     1861 cttgatccgc cttttttgat gtaggtttta acgcggcaaa acatattaat ttttctgatt
     1921 tggaaaaata aagagttagt tctgccccga gcggccgcga ttatcaaaaa ggatcttcac
     1981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2161 accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt
     2221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2461 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtcatga ccaaaatccc
     3061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3241 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3781 gttatcccct gattctgtgg ataaccgtgc ggccgccaat tctaattgta tcgagatcac
     3841 ttttcgtgat ccgctaatca gcgacggtca cattaggttt gccaagtcag ggtatgaacc
     3901 atacgatcag ttttcgtgaa cctggtacgt atattgtggc gtttgtgtat attttcattc
     3961 tttgacaaca atcaatacca acctcaaata ggaaaagtaa taagtttggc gttacacccc
     4021 aaaagacgcc aaacggatcg aacttactca atagcaatta gcgagacaaa acctacgtta
     4081 agacctgtaa ccgatttatc aaagcactct gcggttcttt cttgggaata ttacctggac
     4141 attttgtgcc ctcaagaaac gaggctctac gagcctgttg gagcccctca gacattagcc
     4201 gccacgaatc aaacttttta cgcgattcgg cccaaatcag tttctcacag atcattcgta
     4261 gagtgaaaaa gcacatcgat tatttgatac ccctttgggt taattactgt tgaggtcttt
     4321 gccgtaagtt ccatcgctg
//
