LOCUS       sequence-482776-        2317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid insert contains the aMFx1 and twinStrep C-terminal tag 
            sequences for use in Combinatorial Golden Gate Assembly (BsaI 
            restriction sites)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2317)
  AUTHORS   Cleaver A, Luo R, Smith OB, Murphy L, Schwessinger B, Brock J
  TITLE     High-throughput optimisation of protein secretion in yeast via an 
            engineered biosensor.
  JOURNAL   Trends Biotechnol. 2025 Apr;43(4):838-867. doi: 
            10.1016/j.tibtech.2024.11.010. Epub 2024 Dec 13.
  PUBMED    39674781
REFERENCE   2  (bases 1 to 2317)
  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"; doi: 
            "10.1016/j.tibtech.2024.11.010"; journalName: "Trends Biotechnol"; 
            date: "2025-04"; volume: "43"; issue: "4"; pages: "838-867"
COMMENT     Sequence Label: twinStrep_aMFx1 (4a)
FEATURES             Location/Qualifiers
     source          1..2317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             100..759
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(166..185)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      780..874
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1007..1595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1496..1515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1748..1770
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1767..1783
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1767..1783
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(join(2024..2047,2048..2083,2084..2107))
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(2196..2213)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2196..2212)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2205..2227)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2314..2317,1..99)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
       61 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
      121 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
      181 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
      241 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
      301 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
      361 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
      421 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
      481 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
      541 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
      601 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
      661 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
      721 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg
      781 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
      841 cggttgccgc cgggcgtttt ttattggtga gaatccaagc actagtaaca acttatatcg
      901 tatgggggga cgatttgaag ccccctaccc atttacgctg acttgacggg acggcggctt
      961 tgttgaatag agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1021 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1081 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1141 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1201 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1261 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1321 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1381 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1441 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1501 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1561 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1621 ggttcctggc cttttgctgg ccttttgctc acatgttctt tccagcgaca tatgccatac
     1681 gccgaactga actccaccta atgtcttgga gattgtcgga tagcgcggaa taggattgtg
     1741 ccctctgagc ggataacaat ttcacacagg aaacagctat gaccatgaca catgcatcca
     1801 ccatcgcaga cttatcatca ctgagcctcc acctagccta gttccagatc gtccaggttt
     1861 tgcaccaact actctccgcc agaattttga cgatgagcaa gttccagtga tcgaccgggt
     1921 ctgtaaacta tgccgtctca ggtctcagcc attaatacat tggctgccct ggctttaatt
     1981 gtaaccaatg ccatgcctca gcgtccgcct cacgcttgga tcctttttca aattgtggat
     2041 gggaccaagc agaaccacca gaaccaccac cagaaccacc acctttttca aattgtggat
     2101 gggaccaaga accaccaccg gattgagacc gatgagacga tgcaggtcag gcggaatggc
     2161 acttccttat tccagatgcg tgcggattat gcatgactgg ccgtcgtttt acaacgtcgt
     2221 gactgggaaa accctggcgc ttgtggtgga tcacagttat tggagacttc cttggccgta
     2281 cgatgggttt tgtttcaggt atcaatgaga cgttgat
//
