LOCUS       sequence-403642-        6052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an IPTG inducible mCherry fused to a blue light responsive
            degradation tag (LOVdeg).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6052)
  AUTHORS   Tague N, Coriano-Ortiz C, Sheets MB, Dunlop MJ
  TITLE     Light-inducible protein degradation in E. coli with the LOVdeg tag.
  JOURNAL   Elife. 2024 Jan 25;12:RP87303. doi: 10.7554/eLife.87303.
  PUBMED    38270583
REFERENCE   2  (bases 1 to 6052)
  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"; journalName: "Elife."; 
            date: "2024-01-25"; volume: "12:RP87303. doi"; pages: " 
            10.7554/eLife.87303"
COMMENT     Sequence Label: pBbS5c-mCherry-AsLOV2*(543)
FEATURES             Location/Qualifiers
     source          1..6052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     366..385
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(452..554)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(626..1708)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1670..1689
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1709..1786)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1992..2011
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(2097..2102,2103..2120,2121..2127)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2135..2151
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2192..2899
                     /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(2337..2355)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2605..2624
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3370..3441
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3457..3484
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4015..4965)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(5729..5823)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5844..6052,1..451))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
       61 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
      121 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
      181 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
      241 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
      301 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
      361 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
      421 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
      481 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
      541 ttacgtgccg atcaacgtct cattttcgcc agatatcgac gtcggtgcct aatgagtgag
      601 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
      661 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
      721 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
      781 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
      841 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
      901 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
      961 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     1021 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     1081 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     1141 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     1201 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     1261 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     1321 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     1381 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     1441 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     1501 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     1561 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     1621 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     1681 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     1741 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     1801 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     1861 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     1921 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     1981 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     2041 atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg tttaggcacc ccaggcttta
     2101 cactttatgc ttccggctcg tataatgtgt ggaattgtga gcggataaca atttcagaat
     2161 tcaaaagatc ttttaagaag gagatataca tatggtgagc aagggcgagg aggataacat
     2221 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     2281 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     2341 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     2401 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2461 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2521 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     2581 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2641 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     2701 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     2761 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2821 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     2881 catggacgag ctgtacaagt cgagcggatc aggtttggct actacacttg aacgtattga
     2941 gaagaacttt gtcattactg acccaagatt gccagataat cccattatat tcgcgtccga
     3001 tagtttcttg cagttgacag aatatagccg tgaagaaatt ttgggaagaa actgtcgctt
     3061 cttacaaggc ccggaaaccg atcgcgcaac cgtccgtaag attcgcgacg ccatcgataa
     3121 tcaaaccgag gttacggtgc aattaattaa ctacacgaaa tccggtaaga agttttggaa
     3181 tgtatttcat ttgcaaccca tgcgtgacca gaaaggagat gtacaatatt tcatcggagt
     3241 tcaactcgac ggtacggagc gccttcgcgg ggcagcggaa cgcgaagccg ttatgttgat
     3301 taagaaaacc gcgttccaaa tcgcggaggc ggcataagga tccaaactcg agtaaggatc
     3361 tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     3421 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     3481 tgcgtttata cctagggtac gggttttgct gcccgcaaac gggctgttct ggtgttgcta
     3541 gtttgttatc agaatcgcag atccggcttc agccggtttg ccggctgaaa gcgctatttc
     3601 ttccagaatt gccatgattt tttccccacg ggaggcgtca ctggctcccg tgttgtcggc
     3661 agctttgatt cgataagcag catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct
     3721 gtaacaagtt gtctcaggtg ttcaatttca tgttctagtt gctttgtttt actggtttca
     3781 cctgttctat taggtgttac atgctgttca tctgttacat tgtcgatctg ttcatggtga
     3841 acagctttga atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg
     3901 ttttcatctg tgcatatgga cagttttccc tttgatatgt aacggtgaac agttgttcta
     3961 cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat
     4021 ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga
     4081 gaacgaacca ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact
     4141 ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag
     4201 gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt
     4261 ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc
     4321 agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt
     4381 acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag
     4441 cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt
     4501 ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga
     4561 cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat
     4621 ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg
     4681 gaaaatctca aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc
     4741 tctggttgct ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc
     4801 gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg
     4861 gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg
     4921 actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc
     4981 taggtgattt taatcactat accaattgag atgggctagt caatgataat tactagtcct
     5041 tttcccgggt gatctgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa
     5101 ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca
     5161 agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag
     5221 ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac
     5281 gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc
     5341 aagctcgggc aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct
     5401 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat
     5461 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc
     5521 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt
     5581 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt
     5641 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta
     5701 cccgtcttag gtcatgacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     5761 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     5821 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     5881 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     5941 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     6001 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact ca
//
