LOCUS       sequence-206250-        5368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rhamnose inducible LacZ expression. Used to test burden response in 
            E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5368)
  AUTHORS   Ceroni F, Boo A, Furini S, Gorochowski TE, Borkowski O, Ladak YN, 
            Awan AR, Gilbert C, Stan GB, Ellis T
  TITLE     Burden-driven feedback control of gene expression.
  JOURNAL   Nat Methods. 2018 Mar 26. pii: nmeth.4635. doi: 10.1038/nmeth.4635.
  PUBMED    29578536
REFERENCE   2  (bases 1 to 5368)
  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 
            Methods. 2018 Mar 26. pii: nmeth.4635. doi: 10.1038/nmeth.4635."
COMMENT     Sequence Label: pD864-LacZ
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      70..99
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      196..784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     685..704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(916..943)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(967..1758,1759..1827))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1590..1609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1828..1918)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1927..1957)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2022..2140
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     RBS             2159..2167
                     /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             2173..5247
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(2193..2210)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2193..2209)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2202..2224)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5279..5297)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5293..5340
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gaattcaagc ttgatatcat tcaggacgag cctcagactc cagcgtaact ggactgcaat
       61 caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaagtctt ctcatgacca
      121 aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa
      181 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      241 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      301 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      361 gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      421 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      481 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      661 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      721 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      781 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      841 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      901 agctgatacc gctcgcgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg
      961 ccctcattac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1081 tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg atttatcagc
     1141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1261 gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1801 ggcgacacgg aaatgttgaa tactcatagc tcctgaaaat ctcgataact caaaaaatac
     1861 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     1921 aagacggtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     1981 atttaaatgg tcagtattga gcgatatcta gagaattcgt ccaccacaat tcagcaaatt
     2041 gtgaacatca tcacgttcat ctttccctgg ttgccaatgg cccattttcc tgtcagtaac
     2101 gagaaggtcg cgaattcagg cgctttttag actggtcgta gagaccatga aattctttta
     2161 aggaggtaaa aaatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     2221 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2281 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2341 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     2401 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     2461 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
     2521 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     2581 gaaggccaga cgcgaattat ttttgatggc gttaactcgg cgtttcatct gtggtgcaac
     2641 gggcgctggg tcggttacgg ccaggacagt cgtttgccgt ctgaatttga cctgagcgca
     2701 tttttacgcg ccggagaaaa ccgcctcgcg gtgatggtgc tgcgctggag tgacggcagt
     2761 tatctggaag atcaggatat gtggcggatg agcggcattt tccgtgacgt ctcgttgctg
     2821 cataaaccga ctacacaaat cagcgatttc catgttgcca ctcgctttaa tgatgatttc
     2881 agccgcgctg tactggaggc tgaagttcag atgtgcggcg agttgcgtga ctacctacgg
     2941 gtaacagttt ctttatggca gggtgaaacg caggtcgcca gcggcaccgc gcctttcggc
     3001 ggtgaaatta tcgatgagcg tggtggttat gccgatcgcg tcacactacg tctgaacgtc
     3061 gaaaacccga aactgtggag cgccgaaatc ccgaatctct atcgtgcggt ggttgaactg
     3121 cacaccgccg acggcacgct gattgaagca gaagcctgcg atgtcggttt ccgcgaggtg
     3181 cggattgaaa atggtctgct gctgctgaac ggcaagccgt tgctgattcg aggcgttaac
     3241 cgtcacgagc atcatcctct gcatggtcag gtcatggatg agcagacgat ggtgcaggat
     3301 atcctgctga tgaagcagaa caactttaac gccgtgcgct gttcgcatta tccgaaccat
     3361 ccgctgtggt acacgctgtg cgaccgctac ggcctgtatg tggtggatga agccaatatt
     3421 gaaacccacg gcatggtgcc aatgaatcgt ctgaccgatg atccgcgctg gctaccggcg
     3481 atgagcgaac gcgtaacgcg aatggtgcag cgcgatcgta atcacccgag tgtgatcatc
     3541 tggtcgctgg ggaatgaatc aggccacggc gctaatcacg acgcgctgta tcgctggatc
     3601 aaatctgtcg atccttcccg cccggtgcag tatgaaggcg gcggagccga caccacggcc
     3661 accgatatta tttgcccgat gtacgcgcgc gtggatgaag accagccctt cccggctgtg
     3721 ccgaaatggt ccatcaaaaa atggctttcg ctacctggag agacgcgccc gctgatcctt
     3781 tgcgaatacg cccacgcgat gggtaacagt cttggcggtt tcgctaaata ctggcaggcg
     3841 tttcgtcagt atccccgttt acagggcggc ttcgtctggg actgggtgga tcagtcgctg
     3901 attaaatatg atgaaaacgg caacccgtgg tcggcttacg gcggtgattt tggcgatacg
     3961 ccgaacgatc gccagttctg tatgaacggt ctggtctttg ccgaccgcac gccgcatcca
     4021 gcgctgacgg aagcaaaaca ccagcagcag tttttccagt tccgtttatc cgggcaaacc
     4081 atcgaagtga ccagcgaata cctgttccgt catagcgata acgagctcct gcactggatg
     4141 gtggcgctgg atggtaagcc gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa
     4201 ggtaaacagt tgattgaact gcctgaacta ccgcagccgg agagcgccgg gcaactctgg
     4261 ctcacagtac gcgtagtgca accgaacgcg accgcatggt cagaagccgg gcacatcagc
     4321 gcctggcagc agtggcgtct ggcggaaaac ctcagtgtga cgctccccgc cgcgtcccac
     4381 gccatcccgc atctgaccac cagcgaaatg gatttttgca tcgagctggg taataagcgt
     4441 tggcaattta accgccagtc aggctttctt tcacagatgt ggattggcga taaaaaacaa
     4501 ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc tggataacga cattggcgta
     4561 agtgaagcga cccgcattga ccctaacgcc tgggtcgaac gctggaaggc ggcgggccat
     4621 taccaggccg aagcagcgtt gttgcagtgc acggcagata cacttgctga tgcggtgctg
     4681 attacgaccg ctcacgcgtg gcagcatcag gggaaaacct tatttatcag ccggaaaacc
     4741 taccggattg atggtagtgg tcaaatggcg attaccgttg atgttgaagt ggcgagcgat
     4801 acaccgcatc cggcgcggat tggcctgaac tgccagctgg cgcaggtagc agagcgggta
     4861 aactggctcg gattagggcc gcaagaaaac tatcccgacc gccttactgc cgcctgtttt
     4921 gaccgctggg atctgccatt gtcagacatg tataccccgt acgtcttccc gagcgaaaac
     4981 ggtctgcgct gcgggacgcg cgaattgaat tatggcccac accagtggcg cggcgacttc
     5041 cagttcaaca tcagccgcta cagtcaacag caactgatgg aaaccagcca tcgccatctg
     5101 ctgcacgcgg aagaaggcac atggctgaat atcgacggtt tccatatggg gattggtggc
     5161 gacgactcct ggagcccgtc agtatcggcg gaattccagc tgagcgccgg tcgctaccat
     5221 taccagttgg tctggtgtca aaaataagct cttcaggtta gaggtctcgc ggccgccacc
     5281 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     5341 ctgaaaggag gaactatatc cgggtaac
//
