LOCUS       sequence-377276-        3499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU1 backbone (for TU #1 for plasmid with 2-6 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_63_TU1of2-6_X2homology
FEATURES             Location/Qualifiers
     source          1..3499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             354..365
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             372..1085
                     /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      1097..1168
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1184..1211
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1632..2423,2424..2492))
                     /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     2255..2274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2493..2597)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2700..3288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3189..3208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgtg agaccgaaag tgaaacgtga tttcatgcgt cattttgaac
      241 attttgtaaa tcttatttaa taatgtgtgc ggcaattcac atttaattta tgaatgtttt
      301 cttaacatcg cggcaactca agaaacggca ggttcggatc ttagctacta gagaaagagg
      361 agaaatacta gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg
      421 aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg
      481 caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt
      541 ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc
      601 atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca
      661 cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg
      721 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc
      781 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac
      841 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc
      901 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag
      961 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc
     1021 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt
     1081 acaaatgacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1141 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     1201 gcctttctgc gtttataggt ctcagctgcc aatgagacga cggggtcatc acggctcatc
     1261 atgcgccaaa caaatgtgtg caatacacgc tcggatgact gcatgatgac cgcactgact
     1321 ggggacagca gatccaccta agcctgtgag agaagcagac acccgacaga tcaaggcagt
     1381 taactagtgc actgcagtac ataaccccta ggtcttcctg agagtgacga aggctaaggt
     1441 cacttcttcg tttcctttat tggggtttcc gtgtagcctt cccctgaata gtgtgggacg
     1501 ttttatgaga agccgtaaga aatagccgag cggccgcgat tatcaaaaag gatcttcacc
     1561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1741 ccatctggcc ccagtgctgc aatgataccg cgggacccac gctcaccggc tccagattta
     1801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1921 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgtcatgac caaaatccct
     2641 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2701 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2761 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2821 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2881 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2941 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3001 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3061 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3121 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3181 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3241 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3301 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3361 ttatcccctg attctgtgga taaccgtgcg gccgccaatg tcaaaagatc ctctcatacc
     3421 atattaagta aattgcctcc atttcttttt cctcgggcag agaaactcgc aggcaacttg
     3481 ctctcgaagt ggtcacgtg
//
