LOCUS       sequence-482779-        2313 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  Plasmid insert contains the aMFx4 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_aMFx4 (4a)
FEATURES             Location/Qualifiers
     source          1..2313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..81
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     78..94
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     78..94
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(503..520)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(503..519)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(512..534)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        621..723
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             724..1383
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(790..809)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1404..1498
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1631..2219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2120..2139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aactccacct aatgtcttgg agattgtcgg atagcgcgga ataggattgt gccctctgag
       61 cggataacaa tttcacacag gaaacagcta tgaccatgac acatgcatcc accatcgcag
      121 acttatcatc actgagcctc cacctagcct acaagcacct ttcgtctcag gtctcagcca
      181 ttaatacatt ggctgccctg gctttaattg taaccaatgc catgcctcag cgtccgcctc
      241 acgcttatac ataggttgcc cgggtttaag ttgaagccag tgccaagctt ccgcgtcggc
      301 ctcccttttg tacatcggct gtcccggttt caattgcaac cagtgccagg cttccgcttc
      361 ggcctcacgt ttatacatcg gttgaccagg ttttaactgc agccagtgcc acgcttctgc
      421 ctccctcttg gattgagacc gatgagacga aggtcaggcg gaatggcact tccttattcc
      481 agatgcgtgc ggattatgca tgactggccg tcgttttaca acgtcgtgac tgggaaaacc
      541 ctggcgcttg tggtggatca cagttattgg agacttcctt ggccgtacga tgggttttgt
      601 ttcaggtatc aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa
      661 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
      721 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
      781 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
      841 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
      901 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
      961 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     1021 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     1081 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     1141 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     1201 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     1261 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     1321 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     1381 taatttgata tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact
     1441 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     1501 aagcactagt aacaacttat atcgtatggg gggacgattt gaagccccct acccatttac
     1561 gctgacttga cgggacggcg gctttgttga atagagcgtc agaccccgta gaaaagatca
     1621 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1681 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1741 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1801 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1861 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1921 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1981 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2041 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2101 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2161 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2221 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2281 tctttccagc gacatatgcc atacgccgaa ctg
//
