LOCUS       sequence-412580-        5007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of native L-cysteine desulfidase in E. coli MG1655.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5007)
  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 5007)
  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: pACYC-yhaM
FEATURES             Location/Qualifiers
     source          1..5007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             69..86
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1423..1441)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1437..1484
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1705..2364)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2279..2298
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2365..2467)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2859..2876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3073..3092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3780..4862)
                     /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     4824..4843
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4863..4940)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4986..5005
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4986..5004
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(5005..5007,1..22)
                     /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)."
ORIGIN
        1 attgtgagcg gataacaatt cccctgtaga aataattttg tttaacttta ataaggagat
       61 ataccatgca tcaccaccat catcacggaa gcagctttga ttcgacttta aatccgttat
      121 ggcagcgtta catcctcgcc gttcaggagg aagtaaaacc ggcgctggga tgtactgaac
      181 cgatttcact ggcgctggcg gcggcggttg ctgcggcaga actggaaggt ccggttgaac
      241 gtgtagaagc ctgggtttcg ccaaatctga tgaagaacgg tctgggcgtc accgttcccg
      301 gcacgggaat ggtggggctg ccgattgcgg cggcgctggg ggcgttaggt ggaaatgcca
      361 acgccgggct ggaagtgctg aaagacgcaa cagcgcaggc aattgccgat gccaaagcac
      421 tgctggcggc ggggaaagtc tccgttaaga tccaggaacc ttgcgatgaa atcctcttct
      481 cacgcgccaa agtctggaac ggtgagaagt gggcgtgtgt caccatcgtc ggcgggcata
      541 ccaacattgt gcatatcgag acgcacgatg gtgtggtgtt tacccagcag gcgtgtgtgg
      601 cagagggcga gcaagagtct ccgctgacgg tgctttccag aacgacgctg gctgagatcc
      661 tgaagttcgt caatgaagtc ccgtttgcgg cgatccgctt tattctcgat tccgcgaagc
      721 taaattgtgc gttatcgcag gaaggtttga gcggtaagtg ggggctgcat attggcgcga
      781 cgctggaaaa acagtgcgag cgcggtttgc tggcgaaaga tctctcttca tccattgtga
      841 ttcgtaccag cgcggcatcc gatgcgcgta tgggcggcgc tacgcttccg gctatgagta
      901 actccggctc gggtaaccag gggattaccg caacaatgcc tgtggtggtg gtagcagaac
      961 acttcggagc ggatgatgaa cggctggcgc gtgcgctgat gctttcgcat ttgagcgcaa
     1021 tttacatcca taaccagtta ccgcgtttgt ctgcgctgtg tgccgcaacg accgcagcaa
     1081 tgggggccgc cgccgggatg gcatggctgg tggatgggcg ttatgaaacc atctcgatgg
     1141 cgatcagcag tatgatcggc gatgtcagcg gcatgatttg cgatggtgcg tcgaacagct
     1201 gcgcgatgaa ggtttcgacc agtgcttcgg ctgcgtggaa agcggtgtta atggcgctgg
     1261 atgataccgc cgtgaccggc aatgaaggga ttgtggcgca tgatgttgag cagtcgattg
     1321 ccaacctgtg tgcgttagca agccattcga tgcagcaaac ggatcggcag attatcgaga
     1381 ttatggcgag caaggccaga taattaacct aggctgctgc caccgctgag caataactag
     1441 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat
     1501 ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga
     1561 cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga
     1621 atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag gcgtttaagg
     1681 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     1741 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     1801 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatag tgaaaacggg
     1861 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     1921 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     1981 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     2041 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     2101 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaact ccggatgagc
     2161 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     2221 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     2281 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     2341 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     2401 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     2461 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
     2521 accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag
     2581 tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc
     2641 cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta
     2701 acggcaaaag caccgccgga catcagcgct agcggagtgt atactggctt actatgttgg
     2761 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     2821 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     2881 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     2941 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     3001 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     3061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3241 gctgtgtgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     3301 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     3361 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     3421 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     3481 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     3541 ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg
     3601 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     3661 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3721 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3841 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3901 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3961 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     4021 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     4081 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     4141 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     4201 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     4261 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     4321 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     4381 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4441 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4501 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4561 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4621 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4681 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4741 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4801 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4861 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     4921 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     4981 gaaattaata cgactcacta tagggga
//
