LOCUS       sequence-424617-        4403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of heterologous L-cysteine desulfidase in E. coli 
            MG1655.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4403)
  AUTHORS   Hayes JA, Lunger AW, Sharma AS, Fernez MT, Carrier RL, Koppes AN, 
            Koppes R, Woolston BM
  TITLE     Engineered bacteria titrate hydrogen sulfide and induce 
            concentration-dependent effects on the host in a gut 
            microphysiological system.
  JOURNAL   Cell Rep. 2023 Dec 26;42(12):113481. doi: 
            10.1016/j.celrep.2023.113481. Epub 2023 Nov 18.
  PUBMED    37980564
REFERENCE   2  (bases 1 to 4403)
  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: "Cell Rep.";
            date: "2023-12-26"; volume: "42(12):113481. doi"; pages: " 
            10.1016/j.celrep.2023.113481. Epub 2023 Nov 18"
COMMENT     Sequence Label: pBbA5c-cdl
FEATURES             Location/Qualifiers
     source          1..4403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(280..374)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(395..1054)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     969..988
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1055..1157)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(1229..2311)
                     /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     2273..2292
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2312..2389)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2595..2614
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(2700..2705,2706..2723,2724..2730)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2738..2754
                     /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)."
     terminator      3748..3819
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3835..3862
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3890..3907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4024..4403,1..166))
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4104..4123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
       61 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
      121 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
      181 taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga
      241 agtcagcccc atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc
      301 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
      361 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
      421 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
      481 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
      541 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
      601 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
      661 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
      721 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
      781 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
      841 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
      901 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
      961 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     1021 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     1081 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     1141 ctcttacgtg ccgatcaacg tctcattttc gccagatatc gacgtcggtg cctaatgagt
     1201 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1261 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1321 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     1381 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     1441 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     1501 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     1561 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     1621 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     1681 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     1741 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     1801 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     1861 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     1921 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     1981 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     2041 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     2101 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     2161 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     2221 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     2281 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     2341 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     2401 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     2461 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     2521 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     2581 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     2641 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgtttaggc accccaggct
     2701 ttacacttta tgcttccggc tcgtataatg tgtggaattg tgagcggata acaatttcag
     2761 aattcaaaag atcttttaag aaggagatat acatatgctg gcgaacagcg ttatcgatct
     2821 gatcggaaat acaccgctgg ttaaaatcaa caacatcaac acctttggca acgaaattta
     2881 tgttaaactt gaagggagca acccggggcg cagcaccaaa gaccgcatcg ctctgaaaat
     2941 gatcgaagaa gcggaaaaag aagggctgat cgataaagat accgtgatca ttgaagcgac
     3001 cagcggcaac accggcatcg gattagcgat gatttgcgcg gtgaaaaatt ataagctgaa
     3061 gatcgtgatg ccggatacca tgagcgtgga acgcatccag ttaatgcgcg cttatggcac
     3121 cgaagtgatt ttaaccgatg ggagctttgg catgaaagcg tgcctggaaa aactggaaga
     3181 attaaaaaaa caggaaaaga aatattttat cccgaatcag tttaccaacg ttaacaaccc
     3241 gaaagcgcac tatgaaacca ccgcggagga aatcctgcgc gatatggata acaaagtgga
     3301 tgtttatatc tgcggcaccg gcaccggcgg cagctttagc ggcaccgcga aaaaactgaa
     3361 agaaaaactg ccgaacatca aaacctaccc cgtggaaccg gctagcagcc cgctgttaag
     3421 caagggctat attgggccgc ataaaatcca ggggatgggc atgagcattg gggggattcc
     3481 ggtggtatac gatgggagcc tggcggatgg catcctggtg tgcgaagatg atgaagcttt
     3541 taaaatgatg cgcgaactgt catttaaaga agggatactg gcgggaatca gcaccggggc
     3601 gacccttaaa gcggcgttag attatagcaa agaaaacgcg aacaaaagcc tgcgcattgt
     3661 ggtgttaagc accgacagcg gcgaaaaata tcttagcagc agccatggac tgtaaggatc
     3721 caaactcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3781 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     3841 ccttcgggtg ggcctttctg cgtttatacc tagggatata ttccgcttcc tcgctcactg
     3901 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
     3961 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     4021 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     4081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     4141 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     4201 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     4261 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4321 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     4381 gagttagtct tgaagtcatg cgc
//
