LOCUS       sequence-274953-        3825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Degradation tag variant at C-terminal of mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3825)
  AUTHORS   Teh MY, Ooi KH, Danny Teo SX, Bin Mansoor ME, Shaun Lim WZ, Tan MH
  TITLE     An Expanded Synthetic Biology Toolkit for Gene Expression Control in
            Acetobacteraceae.
  JOURNAL   ACS Synth Biol. 2019 Apr 19;8(4):708-723. doi: 
            10.1021/acssynbio.8b00168. Epub 2019 Mar 22.
  PUBMED    30865830
REFERENCE   2  (bases 1 to 3825)
  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: 
            "10.1021/acssynbio.8b00168"; journalName: "ACS Synth Biol"; date: 
            "2019-04-19- 19"; volume: "8"; issue: "4"; pages: "708-723"
COMMENT     Sequence Label: MYMH 549
FEATURES             Location/Qualifiers
     source          1..3825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(197..283)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     298..320
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        328..362
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     RBS             370..381
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             388..1095
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(533..551)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     801..820
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1102..1134
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AANDENYALAA"
     terminator      1155..1226
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1242..1269
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1276..1296
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1301..1395
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1443..1545
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1546..2205
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1612..1631)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            2354..2462
                     /note="incP origin of transfer"
     CDS             complement(2473..3135)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      join(3136..3825,1..82)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 tcgtgggcag gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg
       61 aagcgagcca gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg
      121 agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc cgtagggggg gcgcgcccag
      181 ctgtctaggg cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
      241 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctt taattaaagc
      301 ggataacaat ttcacacagg agaattcttg acagctagct cagtcctagg tattgtgcta
      361 gctctagaga aagaggagaa atactagatg gtgagcaagg gcgaggagga taacatggcc
      421 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      481 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      541 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
      601 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
      661 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
      721 gtgacccagg actcctcctt gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
      781 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
      841 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
      901 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
      961 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     1021 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1081 gacgagctgt acaagggatc cgctgctaac gacgaaaact acgctctggc tgcttaacct
     1141 gcagggccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1201 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     1261 ctttctgcgt ttatatacta gtagcggccg ctgcagcttg gactcctgtt gatagatcca
     1321 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     1381 tttattggtg agaatccagg ggtccccaat aattacgatt taaattggcg aaaatgagac
     1441 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     1501 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1561 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1621 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1681 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1741 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     1801 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1861 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1921 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1981 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2041 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2101 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     2161 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga cttttgtcct
     2221 tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct
     2281 ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc
     2341 aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct
     2401 tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc
     2461 cgtaggccgg ccctaccggc gcggcagcgt tacccgtgtc ggcggctcca acggctcgcc
     2521 atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc gccgttccca
     2581 ttcctccgtt tcggtcaagg ctggcaggtc tggttccatg cccggaatgc cgggctggct
     2641 gggcggctcc tcgccggggc cggtcggtag ttgctgctcg cccggataca gggtcgggat
     2701 gcggcgcagg tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat caaccaccac
     2761 ggcggcactg aacaccgaca ggcgcaactg gtcgcggggc tggccccacg ccacgcggtc
     2821 attgaccacg taggccgaca cggtgccggg gccgttgagc ttcacgacgg agatccagcg
     2881 ctcggccacc aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc cgatgagctt
     2941 ggaaagtgtc ttctggctga ccaccacggc gttctggtgg cccatctgcg ccacgaggtg
     3001 atgcagcagc attgccgccg tgggtttcct cgcaataagc ccggcccacg cctcatgcgc
     3061 tttgcgttcc gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc cgatttctct
     3121 ggactgcgtg gccatgctta tctccatgcg gtaggggtgc cgcacggttg cggcaccatg
     3181 cgcaatcagc tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca
     3241 gtcaattaca gattttcttt aacctacgca atgagctatt gcggggggtg ccgcaatgag
     3301 ctgttgcgta cccccctttt ttaagttgtt gatttttaag tctttcgcat ttcgccctat
     3361 atctagttct ttggtgccca aagaagggca cccctgcggg gttcccccac gccttcggcg
     3421 cggctccccc tccggcaaaa agtggcccct ccggggcttg ttgatcgact gcgcggcctt
     3481 cggccttgcc caaggtggcg ctgccccctt ggaacccccg cactcgccgc cgtgaggctc
     3541 ggggggcagg cgggcgggct tcgcccttcg actgccccca ctcgcatagg cttgggtcgt
     3601 tccaggcgcg tcaaggccaa gccgctgcgc ggtcgctgcg cgagccttga cccgccttcc
     3661 acttggtgtc caaccggcaa gcgaagcgcg caggccgcag gccggaggct tttccccaga
     3721 gaaaattaaa aaaattgatg gggcaaggcc gcaggccgcg cagttggagc cggtgggtat
     3781 gtggtcgaag gctgggtagc cggtgggcaa tccctgtggt caagc
//
