LOCUS       sequence-384684-        4244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multi Backbone with HO homology arms for Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4244)
  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 4244)
  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_31_backbone_HO_integration
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(630..1218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(710..729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1432..1451)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1443..2258
                     /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             1443..2258
                     /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     complement(1516..1535)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(3053..3080)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3096..3167)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3179..3892)
                     /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"
     RBS             3899..3910
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 agcgatggaa cttacggcaa agacctcaac agtaattaac ccaaaggggt atcaaataat
       61 cgatgtgctt tttcactcta cgaatgatct gtgagaaact gatttgggcc gaatcgcgta
      121 aaaagtttga ttcgtggcgg ctaatgtctg aggggctcca acaggctcgt agagcctcgt
      181 ttcttgaggg cacaaaatgt ccaggtaata ttcccaagaa agaaccgcag agtgctttga
      241 taaatcggtt acaggtctta acgtaggttt tgtctcgcta attgctattg agtaagttcg
      301 atccgtttgg cgtcttttgg ggtgtaacgc caaacttatt acttttccta tttgaggttg
      361 gtattgattg ttgtcaaaga atgaaaatat acacaaacgc cacaatatac gtaccaggtt
      421 cacgaaaact gatcgtatgg ttcataccct gacttggcaa acctaatgtg accgtcgctg
      481 attagcggat cacgaaaagt gatctcgata caattagaat tggcggccgc acggttatcc
      541 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      601 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      661 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      721 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      781 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      841 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      901 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      961 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1021 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1081 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1141 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1201 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1261 aacgaaaact cacgttaagg gattttggtc atgactgatc cttcaactca gcaaaagttc
     1321 gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata
     1381 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1441 ttatgagcca tattcaacgg gaaacgtctt gctccaggcc gcgattaaat tccaacatgg
     1501 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     1561 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1621 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     1681 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     1741 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     1801 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     1861 cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg aataacggtt
     1921 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     1981 aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     2041 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     2101 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     2161 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     2221 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     2281 aacacgcggc cgctcggggc agaactaact ctttattttt ccaaatcaga aaaattaata
     2341 tgttttgccg cgttaaaacc tacatcaaaa aaggcggatc aagatgtatg aaagaaagtg
     2401 cgtagaataa cgaacattca tagctgttct ggaggcttta caaaaggtaa tctttgttag
     2461 gtgcgatttt atccgaaaag caattactct ctatgttagt catacaaact gacttctgtg
     2521 agagaattat tttcatatca acgtaagatc acatggttcc tttatcaagt actactatca
     2581 ttccattata tgacctattt acttcttgaa tcttagagct cataattcaa gcaagttgcg
     2641 gagctaagaa tttcacatgt tgttgaactt aacaatcttc attataccca atcgctgcgt
     2701 gctggaatta tgttaaaagt tacatccttt ttttcatttt tccctacgct cagggcactg
     2761 tactgcccgt gcctgcgatg agatacatca atttaaagcc gcagatgcaa cacgataact
     2821 ctcaggaaga cctaggggtt atgtactgca gtgcactagt ggttcgtaac atctctgtaa
     2881 ctgcttgaga cgtggactgt tgtgatatgc catatatgca gtggtgttcg ccggcgctgg
     2941 ttccggctgt cttgcttagt tgtgagtcgc caggcagtca tcggtatgat ctgtacatga
     3001 ttcgtcagtg tgagcaccac tgacgagcag atttccagct gagacctata aacgcagaaa
     3061 ggcccacccg aaggtgagcc agtgtgactc tagtagagag cgttcaccga caaacaacag
     3121 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctggtcatt
     3181 tgtacagttc atccatacca tgcgtgatgc ccgctgcggt tacgaactcc agcagaacca
     3241 tatgatcgcg tttctcgttc ggatctttag acagaacgct ttgcgtgctc agatagtgat
     3301 tgtctggcag cagaacagga ccatcaccga ttggagtgtt ttgctggtag tgatcagcca
     3361 gctgcacgct gccatcctcc acgttgtggc gaattttaaa attcgcttta atgccatttt
     3421 tttgtttatc ggcggtgatg taaacattgt ggctgttaaa attgtattcc agcttatggc
     3481 ccaggatatt gccgtcctct ttaaagtcaa tgcctttcag ctcaatgcgg tttaccaggg
     3541 tatcgccttc aaatttcact tccgcacgcg ttttgtacgt gccgtcatcc ttaaaggaaa
     3601 tcgtgcgttc ctgcacatag ccttccggca tggcggactt gaagaagtca tgctgcttca
     3661 tatggtccgg ataacgagca aagcactgaa caccataagt cagcgtcgtt accagagtcg
     3721 gccaaggaac cggcagttta ccagtagtac agatgaactt cagcgtcagt ttaccattag
     3781 ttgcgtcacc ttcaccctcg ccacgcacgg aaaacttatg accgttgaca tcaccatcca
     3841 gttccaccag aatagggacg acaccagtga acagctcttc gcctttacgc atctagtatt
     3901 tctcctcttt ctctagtagc taagatccga acctgccgtt tcttgagttg ccgcgatgtt
     3961 aagaaaacat tcataaatta aatgtgaatt gccgcacaca ttattaaata agatttacaa
     4021 aatgttcaaa atgacgcatg aaatcacgtt tcactttcgg tctcacgttg tattgcgacg
     4081 aattgccacg ttgtcgcagt gtctgtactg tgggttaatg tctgtgatcg ccgtcgtgag
     4141 ttcgtgtcat ccgcgcatga ttcagatggt cctggagatc gttgcttgta ccggctgttt
     4201 gtggctctac cacgtctcat cagtctagat gcgaattcag ggac
//
