LOCUS       sequence-410133-        4111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of L-cysteine desulfidase transcriptional regulator 
            in E. coli MG1655.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4111)
  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 4111)
  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-decR
FEATURES             Location/Qualifiers
     source          1..4111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..49)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(720..1802)
                     /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     1764..1783
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1803..1880)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1926..1945
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1926..1944
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1945..1969
                     /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)."
     primer_bind     complement(2491..2509)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2505..2552
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2773..3432)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3347..3366
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3433..3535)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3927..3944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4061..4111,1..495))
                     /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.
                     "
ORIGIN
        1 tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc
       61 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
      121 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
      181 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
      241 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
      301 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
      361 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
      421 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
      481 ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg
      541 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
      601 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
      661 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
      721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      781 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
      841 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
      901 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
      961 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     1021 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     1081 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     1141 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     1201 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     1261 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     1321 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     1381 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     1441 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     1501 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     1561 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     1621 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     1681 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     1741 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     1801 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     1861 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     1921 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctgtagaaat
     1981 aattttgttt aactttaata aggagatata ccatgttaga taaaattgac cgtaagctgc
     2041 tggccttact gcagcaggat tgcaccctct ctttgcaggc actggctgaa gccgttaatc
     2101 tgacaaccac cccttgctgg aagcgcctga aacggctgga ggacgacggt atccttatcg
     2161 gcaaagtcgc cctgctggat ccggaaaaaa taggcctcgg cctgaccgct tttgtgctga
     2221 taaaaacgca acatcacagc agcgaatggt attgccgctt tgtcacggtg gttaccgaaa
     2281 tgccagaagt gctggggttc tggcgcatgg ctggtgaata cgattatctg atgcgcgtcc
     2341 aggttgccga catgaaacgc tacgacgagt tttataagcg tctggtaaac agcgtgccgg
     2401 ggctgtcgga cgtcacttcc agcttcgcga tggaacagat taaatacacc acttctttac
     2461 ccatcgaata attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     2521 gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca
     2581 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     2641 tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt
     2701 catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact
     2761 gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     2821 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     2881 cagcaccttg tcgccttgcg tataatattt gcccatagtg aaaacggggg cgaagaagtt
     2941 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     3001 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     3061 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     3121 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     3181 tatcaccagc tcaccgtctt tcattgccat acggaactcc ggatgagcat tcatcaggcg
     3241 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     3301 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     3361 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     3421 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     3481 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc
     3541 tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac caggatttat
     3601 ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga
     3661 tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg
     3721 tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca
     3781 ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg
     3841 tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat
     3901 atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     3961 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     4021 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     4081 ctgacaagca tcacgaaatc tgacgctcaa a
//
