LOCUS       sequence-384685-        3404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multi Backbone with X2 homology arms for Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3404)
  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 3404)
  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_30_backbone_X2_integration
FEATURES             Location/Qualifiers
     source          1..3404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             334..345
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             352..1065
                     /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      1077..1148
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1164..1191
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1566..2381)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1566..2381)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2289..2308
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2373..2392
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2606..3194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3095..3114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccctgaatt cgcatctaga ctgatgagac gtggtagagc cacaaacagc cggtacaagc
       61 aacgatctcc aggaccatct gaatcatgcg cggatgacac gaactcacga cggcgatcac
      121 agacattaac ccacagtaca gacactgcga caacgtggca attcgtcgca atacaacgtg
      181 agaccgaaag tgaaacgtga tttcatgcgt cattttgaac attttgtaaa tcttatttaa
      241 taatgtgtgc ggcaattcac atttaattta tgaatgtttt cttaacatcg cggcaactca
      301 agaaacggca ggttcggatc ttagctacta gagaaagagg agaaatacta gatgcgtaaa
      361 ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac
      421 ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg
      481 ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct ggtaacgacg
      541 ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc
      601 ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac
      661 ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt
      721 gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa gctggaatac
      781 aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
      841 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag
      901 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
      961 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
     1021 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc aggcatcaaa
     1081 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1141 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttataggt
     1201 ctcagctgga aatctgctcg tcagtggtgc tcacactgac gaatcatgta cagatcatac
     1261 cgatgactgc ctggcgactc acaactaagc aagacagccg gaaccagcgc cggcgaacac
     1321 cactgcatat atggcatatc acaacagtcc acgtctcaag cagttacaga gatgttacga
     1381 accactagtg cactgcagta cataacccct aggtcttcct gagagtgacg aaggctaagg
     1441 tcacttcttc gtttccttta ttggggtttc cgtgtagcct tcccctgaat agtgtgggac
     1501 gttttatgag aagccgtaag aaatagccga gcggccgcgt gttacaacca attaaccaat
     1561 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1621 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1681 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1741 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1801 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     1861 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1921 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1981 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     2041 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     2101 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     2161 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2221 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2281 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2341 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     2401 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     2461 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     2521 aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg
     3301 ccaatgtcaa aagatcctct cataccatat taagtaaatt gcctccattt ctttttcctc
     3361 gggcagagaa actcgcaggc aacttgctct cgaagtggtc acgt
//
