LOCUS       sequence-478389-        2313 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Plasmid insert contains the aMFx1 C-terminal tag sequence with the 
            TEV cleavage site 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 Jul 14, 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_TEV_aMFx1 (4a)
FEATURES             Location/Qualifiers
     source          1..2313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             70..90
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(357..373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(357..373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(370..392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(545..1133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(625..644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1266..1360)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1381..2040)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1955..1974
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2041..2143)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2230..2252
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2244..2261
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2245..2261
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aatacgtagc gtggtcgcat aaaagttaat tctacgggac taaagaggcc gtctcatcgg
       61 tctcaatccg aaaatttgta cttccaatcc tggcattggt tacaattaaa gccagggcag
      121 ccaatgtatt aatggctgag acctgagacg aaaggtgctt gtgttccgac aggctaggat
      181 ataatcctaa ggcgttaccc caatcgttca gcgtgggatt tgctacaact cctgaacgct
      241 acatgtacga aaccatgtta tgtatgcaca aggccaacaa taggacgtag ccttgaagtt
      301 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
      361 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
      421 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
      481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1141 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
     1201 agcgtaaatg ggtagggggc ttcaaatcgt ccccccatac gatataagtt gttactagtg
     1261 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     1321 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     1381 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     1441 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     1501 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     1561 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     1621 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     1681 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     1741 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     1801 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     1861 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     1921 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     1981 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     2041 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     2101 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attgatacct
     2161 gaaacaaaac ccatcgtacg gccaaggaag tctccaataa ctgtgatcca ccacaagcgc
     2221 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tcatgcataa tccgcacgca
     2281 tctggaataa ggaagtgcca ttccgcctga cct
//
