LOCUS       sequence-410114-        4918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of L-cysteine importer in E. coli MG1655.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4918)
  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 4918)
  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: pMF2-yhaO
FEATURES             Location/Qualifiers
     source          1..4918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..37)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     178..200
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(194..385)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      496..541
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(557..1435)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1462..1746
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1669..1688
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(3204..3995)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3996..4087)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4096..4684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4585..4604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4838..4855
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(4870..4918,1..92)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
       61 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgcaaaca cccgctgacg
      121 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      181 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
      241 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
      301 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
      361 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
      421 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
      481 aagaggacat ccggtcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
      541 agacttaggg accctcttat gacaacttga cggctacatc attcactttt tcttcacaac
      601 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
      661 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
      721 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
      781 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
      841 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
      901 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
      961 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     1021 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     1081 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     1141 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     1201 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     1261 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     1321 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     1381 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     1441 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
     1501 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
     1561 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
     1621 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
     1681 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
     1741 ctccataccc gtttttttgg gctagcgaat ttgagctaag gaggaaaaaa aaatggaaat
     1801 tgcatcgaat aaaggcgtca ttgcagacgc ttcgaccccg gcgggtcgtg ctggaatgag
     1861 tgagagcgag tggcgagaag cgatcaaatt cgacagtact gacaccggct gggtgattat
     1921 gagtatcggg atggcgattg gcgcggggat tgtttttctc ccggtgcagg tcggtttgat
     1981 gggattgtgg gtatttttgc tctcatcggt gattggttac ccggcaatgt atctgtttca
     2041 gcgattgttt attaatacgc tggcagaatc accagaatgt aaagattacc cgagcgtcat
     2101 tagcggttat ttaggtaaaa actggggcat cctgttaggt gcgctctatt tcgtaatgct
     2161 ggtgatttgg atgttcgtct attccaccgc catcaccaac gatagtgctt cctacctgca
     2221 taccttcggc gtgacggaag ggttgctgtc agacagtccc ttttatggtc tggtactgat
     2281 ttgcattctg gtggcgatct cctcacgcgg cgagaaattg ttattcaaaa tttcgaccgg
     2341 catggtgctg accaagctgc tggtggtcgc ggcgctgggc gtgtcgatgg tgggaatgtg
     2401 gcatctgtat aacgtcggtt cgctaccgcc gctggggctg ctggtgaaaa acgccattat
     2461 tacgctgccg tttaccctga cgtcgattct gtttatccag acgttaagtc cgatggtgat
     2521 ctcttatcgc tcgcgggaaa aatcaattga agtggcgcgg cataaagcat tgcgggcaat
     2581 gaatatcgcg tttggcattt tgtttgtcac cgtctttttc tacgccgtgt cgttcacgct
     2641 ggcgatggga catgacgaag cggtaaaagc ctatgagcag aatatttccg cgctggcgat
     2701 tgccgcacag tttattagcg gtgacggcgc agcgtgggtg aaagtggtca gcgtcattct
     2761 caatatcttt gctgtcatga ctgcattctt tggcgtctat ttaggctttc gcgaagcaac
     2821 gcaagggatc gtaatgaaca tcctgcgtcg caagatgcct gccgagaaga ttaacgaaaa
     2881 tctcgttcag cgcggcatca tgattttcgc cattttgctg gcctggagcg ccatcgtact
     2941 gaacgcaccg gtgttgagct tcacctctat ctgtagcccg attttcggca tggtagggtg
     3001 cctgatcccg gcgtggctgg tttacaaagt accagcattg cacaaataca aagggatgtc
     3061 tctgtacctg attatcgtca ctggtttgtt gctgtgtgtt tctccgttcc tggcattttc
     3121 ttgaacttaa ttaacggcac tcctcagcaa atataatgac cctcttgata acccaagagg
     3181 gcatttttta atgcccatgg cgtttatttg ccgactacct tggtgatctc gcctttcacg
     3241 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
     3301 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     3361 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     3421 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     3481 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     3541 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     3601 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     3661 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     3721 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     3781 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     3841 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     3901 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     3961 tgagtcgata cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga
     4021 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4081 aggccaaata ggccgttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4141 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccac ctctttttcc
     4201 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     4261 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4681 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4741 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4801 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4861 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgc
//
