LOCUS       sequence-410139-        4600 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 4600)
  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 4600)
  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-cdl
FEATURES             Location/Qualifiers
     source          1..4600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             21..38
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(985..1003)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      999..1046
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1267..1926)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1841..1860
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1927..2029)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2421..2438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2555..3100)
                     /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(2635..2654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3325..4407)
                     /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     4369..4388
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4408..4485)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4531..4550
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4531..4549
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4550..4574
                     /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 taataaggag atataccatg catcaccacc atcatcacgg aagcagcctg gcgaacagcg
       61 ttatcgatct gatcggaaat acaccgctgg ttaaaatcaa caacatcaac acctttggca
      121 acgaaattta tgttaaactt gaagggagca acccggggcg cagcaccaaa gaccgcatcg
      181 ctctgaaaat gatcgaagaa gcggaaaaag aagggctgat cgataaagat accgtgatca
      241 ttgaagcgac cagcggcaac accggcatcg gattagcgat gatttgcgcg gtgaaaaatt
      301 ataagctgaa gatcgtgatg ccggatacca tgagcgtgga acgcatccag ttaatgcgcg
      361 cttatggcac cgaagtgatt ttaaccgatg ggagctttgg catgaaagcg tgcctggaaa
      421 aactggaaga attaaaaaaa caggaaaaga aatattttat cccgaatcag tttaccaacg
      481 ttaacaaccc gaaagcgcac tatgaaacca ccgcggagga aatcctgcgc gatatggata
      541 acaaagtgga tgtttatatc tgcggcaccg gcaccggcgg cagctttagc ggcaccgcga
      601 aaaaactgaa agaaaaactg ccgaacatca aaacctaccc cgtggaaccg gctagcagcc
      661 cgctgttaag caagggctat attgggccgc ataaaatcca ggggatgggc atgagcattg
      721 gggggattcc ggtggtatac gatgggagcc tggcggatgg catcctggtg tgcgaagatg
      781 atgaagcttt taaaatgatg cgcgaactgt catttaaaga agggatactg gcgggaatca
      841 gcaccggggc gacccttaaa gcggcgttag attatagcaa agaaaacgcg aacaaaagcc
      901 tgcgcattgt ggtgttaagc accgacagcg gcgaaaaata tcttagcagc agccatggac
      961 tgtaattaac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     1021 ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
     1081 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     1141 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     1201 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     1261 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     1321 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1381 cttgtcgcct tgcgtataat atttgcccat agtgaaaacg ggggcgaaga agttgtccat
     1441 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     1501 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1561 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     1621 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1681 cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag
     1741 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1801 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1861 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1921 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     1981 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     2041 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     2101 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     2161 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     2221 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     2281 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     2341 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     2401 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     2461 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     2521 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     2581 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     2641 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     2701 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     2761 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     2821 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     2881 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     2941 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     3001 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     3061 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     3121 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     3181 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
     3241 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     3301 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3361 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     3421 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     3481 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     3541 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     3601 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     3661 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     3721 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     3781 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     3841 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     3901 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     3961 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     4021 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     4081 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     4141 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     4201 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     4261 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     4321 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     4381 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     4441 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     4501 ctcccttatg cgactcctgc attaggaaat taatacgact cactataggg gaattgtgag
     4561 cggataacaa ttcccctgta gaaataattt tgtttaactt
//
