LOCUS       sequence-383070-        3499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU4 backbone (for TU #4 for plasmid with 5-6 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_69_TU4of5-6_X2homology
FEATURES             Location/Qualifiers
     source          1..3499
                     /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(1633..2424,2425..2493))
                     /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     2256..2275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2494..2598)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2701..3289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3190..3209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccctgaatt 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 tctcagctgg gtatgagacg gcctgcacta gacgaactag
     1261 gcaagatgcg tccaatccgt ctaaacatgg tgacaacgct ggacagatga cgtaacaccg
     1321 agccacatcc tgaaatcgag gcaggctaac cgaaaccgtg acaatgcaaa gagacagcct
     1381 gacactagtg cactgcagta cataacccct aggtcttcct gagagtgacg aaggctaagg
     1441 tcacttcttc gtttccttta ttggggtttc cgtgtagcct tcccctgaat agtgtgggac
     1501 gttttatgag aagccgtaag aaatagccga gcggccgcga ttatcaaaaa ggatcttcac
     1561 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1741 accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt
     1801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2521 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2581 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtcatga ccaaaatccc
     2641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2821 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2941 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3001 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3061 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3361 gttatcccct gattctgtgg ataaccgtgc ggccgccaat gtcaaaagat cctctcatac
     3421 catattaagt aaattgcctc catttctttt tcctcgggca gagaaactcg caggcaactt
     3481 gctctcgaag tggtcacgt
//
