LOCUS       sequence-406411-        5968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables dual growth and fluorescence based enrichment of RamR 
            biosensors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5968)
  AUTHORS   d'Oelsnitz S, Kim W, Burkholder NT, Javanmardi K, Thyer R, Zhang Y, 
            Alper HS, Ellington AD
  TITLE     Using fungible biosensors to evolve improved alkaloid biosyntheses.
  JOURNAL   Nat Chem Biol. 2022 Jul 7. pii: 10.1038/s41589-022-01072-w. doi: 
            10.1038/s41589-022-01072-w.
  PUBMED    35799063
REFERENCE   2  (bases 1 to 5968)
  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: "Nat Chem 
            Biol. 2022 Jul 7. pii: 10.1038/s41589-022-01072-w. doi: 
            10.1038/s41589-022-01072-w."
COMMENT     Sequence Label: pSELIS-RamR
FEATURES             Location/Qualifiers
     source          1..5968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      79..624
                     /direction=RIGHT
                     /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     525..544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     741..758
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1150..1252
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1253..1912
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISRWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1319..1338)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2824..3534
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETF"
     RBS             3726..3734
                     /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             4464..4481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4496..4567
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4583..4610
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4662..4739
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4740..5822
                     /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="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKAEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGLLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDHLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4759..4778)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gacttcaggt gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg
       61 attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
      121 cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa
      181 ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta
      241 accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg
      301 gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc
      361 ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg
      421 aactgagtgt taggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta
      481 aaccgaaagg caggaacagg agagcgcacg agggagccac cagggggaaa cgcctggtat
      541 ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg
      601 tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa
      661 gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc
      721 gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc
      781 acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact
      841 gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgatgtccg
      901 gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg acagctgata
      961 gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca gtaagttggc
     1021 agcatcaccc gacgcacttt gcgccgaata aatacctgtg acggaagatc acttcgcaga
     1081 ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact tttggcgaaa
     1141 atgagacgtt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
     1201 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
     1261 aaaaatcact ggatatacca ccgttgatat atcccgatgg catcgtaaag aacattttga
     1321 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1381 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1441 tgcccgcctg atgaatgctc atccggagtt ccgtatggca atgaaagacg gtgagctggt
     1501 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     1561 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     1621 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     1681 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1741 ggacaacttc ttcgcccccg ttttcactat gggcaaatat tatacgcaag gcgacaaggt
     1801 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     1861 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
     1921 aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg ataataagcg
     1981 gatgaatggc agaaattcga aagcaaattc gacccggtcg tcggttcagg gcagggtcgt
     2041 taaatagccg cttatgtcta ttgctggttt accggtttat tgactaccgg aagcagtgtg
     2101 accgtgtgct tctcaaatgc ctgaggtttc agaaataata aaaaagccgg attaataatc
     2161 tggcttttta tattctctct ttattaatct tgttcttcag caacaaaatg aacgcaatta
     2221 ccagctggat ctctcaaagc aaattctcta ccccaaggtt gttcaccaat ttcggtcata
     2281 gctggaccag aagcatctct aaaattggta gaaacaactt cggaccattc ggcatataat
     2341 tcatccaaac ctctaaccca aacccaagcc aaagtattat ctggaacaac ttgatcttga
     2401 acggcggaaa tgaacaaggt aacatcatct ctaacaacac cagcgaaatc atcttcaacg
     2461 aaatctctgg agaaacccaa tctatcagtc caaaattcaa cagcaccagc aacatctcta
     2521 gcggtcaaaa ctggaacagc agatgtcaat ttagacatat gtcaacctcc tccaataacc
     2581 gtcactgctg acgactctac cattatcact tgcaaccatt atcaccgcca gaggtaaaat
     2641 agtcaacacg cacggtgtta gtcttttcgc cggtgtattt cttgcaaaaa aaaaccccgc
     2701 cgaagcgggg tttttttttt acagggctat aaggtagatc cgggagctgt tgacggcgta
     2761 tctttgcttt ctataatgag tgcttactca ctcataatca agtactagag aaggagatct
     2821 agaatgagca caaaaaagaa accattaaca caagagcagc ttgaggacgc acgtcgcctt
     2881 aaagcaattt atgaaaaaaa gaaaaatgaa cttggcttat cccaggaatc tgtcgcagac
     2941 aagatgggga tggggcagtc aggcgttggt gctttattta atggcatcaa tgcattaaat
     3001 gcttataacg ccgcattgct tgcaaaaatt ctcaaagtta gcgttgaaga atttagccct
     3061 tcaatcgcca gagaaatcta cgagatgtat gaagcggtta gtatgcagcc gtcacttaga
     3121 agtgagtatg agtaccctgt tttttctcat gttcaggcag ggatgttctc acctgagctt
     3181 agaaccttta ccaaaggtga tgcggagaga tgggtaagca caaccaaaaa agccagtgat
     3241 tctgcattct ggcttgaggt tgaaggtaat tccatgaccg caccaacagg ctccaagcca
     3301 agctttcctg acggaatgtt aattctcgtt gaccctgagc aggctgttga gccaggtgat
     3361 ttctgcatag ccagacttgg gggtgatgag tttaccttca agaaactgat cagggatagc
     3421 ggtcaggtgt ttttacaacc actaaaccca cagtacccaa tgatcccatg caatgagagt
     3481 tgttccgttg tggggaaagt tatcgctagt cagtggcctg aagagacgtt ttaacgataa
     3541 tgtcgggcaa tcaggtgcga ctcggtacca aattccagaa aagaggggag cgggaaaccg
     3601 ctcccctttt ttcgttttgg tcccaatcta tcgattgtat gggctgtcga cctttgaaaa
     3661 gtaccttgac ggcgtatctt tgctttctat aatgagtgct tactcactca tagggtttga
     3721 tttaataagg aggtatacat atggcatcca agggcgagga gctctttact ggcgtagtac
     3781 caattctcgt agagctcgat ggcgatgtaa atggccataa gttttccgta cgcggcgagg
     3841 gcgagggcga tgcaactaac ggcaagctca ctctcaagtt tatttgtact actggcaagc
     3901 tcccagtacc atggccaact ctcgtaacta ctctgaccta tggcgtacaa tgtttttccc
     3961 gctatccaga tcacatgaag caacatgatt tttttaagtc cgcaatgcca gagggctatg
     4021 tacaagagcg cactattagc tttaaggatg atggcaccta taagactcgc gcagaggtaa
     4081 agtttgaggg cgatactctc gtaaatcgca ttgagctcaa gggcattgat tttaaggagg
     4141 atggcaatat tctcggccat aagctggagt ataatttcaa ttcccataat gtatatatta
     4201 ccgcagataa gcaaaagaat ggcattaagg cgaattttaa gattcgccat aatgtggagg
     4261 atggctccgt acaactcgca gatcattatc aacaaaatac tccaattggc gatggcccag
     4321 tactcctccc agataatcat tatctctcca ctcaatccgt gctctccaaa gatccaaatg
     4381 agaagcgcga tcacatggta ctcctggagt ttgtaactgc agcaggcatt actcatggca
     4441 tggatgagct ctataagctc gagcaccacc accaccacca ctaataacca ggcatcaaat
     4501 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     4561 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataattt
     4621 cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca
     4681 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     4741 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     4801 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gcggaagcgg
     4861 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     4921 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     4981 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     5041 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     5101 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     5161 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     5221 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     5281 aaatcgcgct gttagcgggc ctattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     5341 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     5401 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     5461 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     5521 ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa gacagctcat
     5581 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     5641 tggaccactt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     5701 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     5761 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5821 gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg
     5881 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactaacatg
     5941 agaattacaa cttatatcgt atggggct
//
