LOCUS       sequence-482780-        2313 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  Plasmid insert contains the aMFx1 C-terminal tag sequence 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 2313)
  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 2313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 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: C-term_aMFx1 (4a)
FEATURES             Location/Qualifiers
     source          1..2313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(285..379)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(400..1059)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     974..993
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1060..1162)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1249..1271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1263..1280
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1264..1280
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1689..1705)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1689..1705)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1702..1724)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(1877..2313,1..152))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1957..1976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
       61 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      121 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
      181 ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca gcgtaaatgg gtagggggct
      241 tcaaatcgtc cccccatacg atataagttg ttactagtgc ttggattctc accaataaaa
      301 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
      361 ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc cctgccactc
      421 atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc
      481 atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc
      541 catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt
      601 gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa
      661 ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg
      721 gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac
      781 ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg
      841 aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
      901 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      961 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
     1021 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
     1081 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     1141 ggaacctctt acgtgccgat caacgtctca ttgatacctg aaacaaaacc catcgtacgg
     1201 ccaaggaagt ctccaataac tgtgatccac cacaagcgcc agggttttcc cagtcacgac
     1261 gttgtaaaac gacggccagt catgcataat ccgcacgcat ctggaataag gaagtgccat
     1321 tccgcctgac ctgttagagg ctcactatag catcaggtag actagcatgt atctttactc
     1381 ccaggggaat acgggtgcgt ggacaaatga gcagcaaacg taaattctcg gcgtgcttgg
     1441 tgtctcgcgt ctcatcggtc tcaatccaag cgtgaggcgg acgctgaggc atggcattgg
     1501 ttacaattaa agccagggca gccaatgtat taatggctga gacctgagac gaaaggtgct
     1561 tgtgttccga caggctagga tataatccta aggcgttacc ccaatcgttc agcgtgggat
     1621 ttgctacaac taaggctagg tggaggctca gtgatgataa gtctgcgatg gtggatgcat
     1681 gtgtcatggt catagctgtt tcctgtgtga aattgttatc cgctcagagg gcacaatcct
     1741 attccgcgct atccgacaat ctccaagaca ttaggtggag ttcagttcgg cgtatggcat
     1801 atgtcgctgg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1861 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1921 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1981 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2041 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2101 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2161 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2221 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2281 tcttgaagtg gtggcctaac tacggctaca cta
//
