LOCUS       sequence-410137-        5703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of native L-cysteine desulfidase and RFP in E. coli 
            MG1655.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5703)
  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 5703)
  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-RFP
FEATURES             Location/Qualifiers
     source          1..5703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..38
                     /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             85..102
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1439..1457)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1453..1500
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1720..2397)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2404..2412
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(2417..3076)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2991..3010
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3077..3179)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3571..3588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3785..3804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4492..5574)
                     /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     5536..5555
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5575..5652)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5698..5703,1..14)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5698..5703,1..13)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gactcactat aggggaattg tgagcggata acaattcccc tgtagaaata attttgttta
       61 actttaataa ggagatatac catgcatcac caccatcatc acggaagcag ctttgattcg
      121 actttaaatc cgttatggca gcgttacatc ctcgccgttc aggaggaagt aaaaccggcg
      181 ctgggatgta ctgaaccgat ttcactggcg ctggcggcgg cggttgctgc ggcagaactg
      241 gaaggtccgg ttgaacgtgt agaagcctgg gtttcgccaa atctgatgaa gaacggtctg
      301 ggcgtcaccg ttcccggcac gggaatggtg gggctgccga ttgcggcggc gctgggggcg
      361 ttaggtggaa atgccaacgc cgggctggaa gtgctgaaag acgcaacagc gcaggcaatt
      421 gccgatgcca aagcactgct ggcggcgggg aaagtctccg ttaagatcca ggaaccttgc
      481 gatgaaatcc tcttctcacg cgccaaagtc tggaacggtg agaagtgggc gtgtgtcacc
      541 atcgtcggcg ggcataccaa cattgtgcat atcgagacgc acgatggtgt ggtgtttacc
      601 cagcaggcgt gtgtggcaga gggcgagcaa gagtctccgc tgacggtgct ttccagaacg
      661 acgctggctg agatcctgaa gttcgtcaat gaagtcccgt ttgcggcgat ccgctttatt
      721 ctcgattccg cgaagctaaa ttgtgcgtta tcgcaggaag gtttgagcgg taagtggggg
      781 ctgcatattg gcgcgacgct ggaaaaacag tgcgagcgcg gtttgctggc gaaagatctc
      841 tcttcatcca ttgtgattcg taccagcgcg gcatccgatg cgcgtatggg cggcgctacg
      901 cttccggcta tgagtaactc cggctcgggt aaccagggga ttaccgcaac aatgcctgtg
      961 gtggtggtag cagaacactt cggagcggat gatgaacggc tggcgcgtgc gctgatgctt
     1021 tcgcatttga gcgcaattta catccataac cagttaccgc gtttgtctgc gctgtgtgcc
     1081 gcaacgaccg cagcaatggg ggccgccgcc gggatggcat ggctggtgga tgggcgttat
     1141 gaaaccatct cgatggcgat cagcagtatg atcggcgatg tcagcggcat gatttgcgat
     1201 ggtgcgtcga acagctgcgc gatgaaggtt tcgaccagtg cttcggctgc gtggaaagcg
     1261 gtgttaatgg cgctggatga taccgccgtg accggcaatg aagggattgt ggcgcatgat
     1321 gttgagcagt cgattgccaa cctgtgtgcg ttagcaagcc attcgatgca gcaaacggat
     1381 cggcagatta tcgagattat ggcgagcaag gccagataat taacctaggc tgctgccacc
     1441 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1501 ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     1561 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     1621 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     1681 aaccaggcgt ttaagggcac caataactgc cttaaaaaat taagcaccgg tggagtgacg
     1741 accttcagca cgttcgtact gttcaacgat ggtgtagtct tcgttgtggg aggtgatgtc
     1801 cagtttgatg tcggttttgt aagcacccgg cagctgaacc ggttttttag ccatgtaggt
     1861 ggttttaact tcagcgtcgt agtgaccacc gtctttcagt ttcagacgca ttttgatttc
     1921 acctttcaga gcaccgtctt ccgggtacat acgttcggtg gaagcttccc aacccatggt
     1981 ttttttctgc ataaccggac cgtcggacgg gaagttggta ccacgcagtt taactttgta
     2041 gatgaactca ccgtcttgca gggaggagtc ctgggtaacg gtaacaacac caccgtcttc
     2101 gaagttcata acacgttccc atttgaaacc ttccgggaag gacagtttca ggtagtccgg
     2161 gatgtcagcc gggtgtttaa cgtaagcttt ggaaccgtac tggaactgcg gggacaggat
     2221 gtcccaagcg aacggcagcg gaccaccttt ggtaactttc agtttagcgg tctgggtacc
     2281 ttcgtacgga cgaccttcac cttcaccttc gatttcgaac tcgtgaccgt taacggaacc
     2341 ttccatacga actttgaaac gcatgaactc tttgataacg tcttcgctac tcgccatgtt
     2401 aatacctcct tattaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     2461 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     2521 gcatcagcac cttgtcgcct tgcgtataat atttgcccat agtgaaaacg ggggcgaaga
     2581 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     2641 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     2701 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     2761 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     2821 cccatatcac cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca
     2881 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     2941 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     3001 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     3061 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     3121 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     3181 cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat
     3241 ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg
     3301 gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct
     3361 tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa
     3421 agcaccgccg gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg
     3481 agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca
     3541 gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     3601 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     3661 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     3721 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     3781 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3841 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3901 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3961 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4021 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     4081 gttagtcttg aagtcacgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     4141 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     4201 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     4261 atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat
     4321 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta
     4381 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     4441 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     4501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4561 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     4621 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     4681 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     4741 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     4801 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     4861 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     4921 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     4981 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     5041 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     5101 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     5161 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     5221 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     5281 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     5341 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     5401 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     5461 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     5521 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     5581 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     5641 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa
     5701 tac
//
