LOCUS       sequence-210619-        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      58..87
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      184..772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     673..692
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(904..931)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(955..1746,1747..1815))
                     /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     1578..1597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1816..1906)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1915..1945)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2010..2128
                     /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             2147..2155
                     /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             2161..5235
                     /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(2181..2198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2181..2197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2190..2212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5267..5285)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5281..5328
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gatatcattc aggacgagcc tcagactcca gcgtaactgg actgcaatca actcactggc
       61 tcaccttcac gggtgggcct ttcttcggta gaagtcttct catgaccaaa atcccttaac
      121 gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      181 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      241 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      301 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca
      361 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      421 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      481 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      541 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      601 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      661 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      721 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      781 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      841 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      901 tcgcgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcattacca
      961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagcgc
     1081 tgcgatgata ccgcgagaac cacgctcacc ggctccggat ttatcagcaa taaaccagcc
     1141 agcaggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1261 tgccatcgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1801 atgttgaata ctcatagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     1861 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagaa gacggtcaaa
     1921 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     1981 agtattgagc gatatctaga gaattcgtcc accacaattc agcaaattgt gaacatcatc
     2041 acgttcatct ttccctggtt gccaatggcc cattttcctg tcagtaacga gaaggtcgcg
     2101 aattcaggcg ctttttagac tggtcgtaga gaccatgaaa ttcttttaag gaggtaaaaa
     2161 atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2221 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2281 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     2341 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     2401 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     2461 tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     2521 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     2581 cgaattattt ttgatggcgt taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc
     2641 ggttacggcc aggacagtcg tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc
     2701 ggagaaaacc gcctcgcggt gatggtgctg cgctggagtg acggcagtta tctggaagat
     2761 caggatatgt ggcggatgag cggcattttc cgtgacgtct cgttgctgca taaaccgact
     2821 acacaaatca gcgatttcca tgttgccact cgctttaatg atgatttcag ccgcgctgta
     2881 ctggaggctg aagttcagat gtgcggcgag ttgcgtgact acctacgggt aacagtttct
     2941 ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc
     3001 gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga aaacccgaaa
     3061 ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca caccgccgac
     3121 ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg gattgaaaat
     3181 ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg tcacgagcat
     3241 catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatat cctgctgatg
     3301 aagcagaaca actttaacgc cgtgcgctgt tcgcattatc cgaaccatcc gctgtggtac
     3361 acgctgtgcg accgctacgg cctgtatgtg gtggatgaag ccaatattga aacccacggc
     3421 atggtgccaa tgaatcgtct gaccgatgat ccgcgctggc taccggcgat gagcgaacgc
     3481 gtaacgcgaa tggtgcagcg cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg
     3541 aatgaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa atctgtcgat
     3601 ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac cgatattatt
     3661 tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc gaaatggtcc
     3721 atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgatcctttg cgaatacgcc
     3781 cacgcgatgg gtaacagtct tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat
     3841 ccccgtttac agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat
     3901 gaaaacggca acccgtggtc ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc
     3961 cagttctgta tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc gctgacggaa
     4021 gcaaaacacc agcagcagtt tttccagttc cgtttatccg ggcaaaccat cgaagtgacc
     4081 agcgaatacc tgttccgtca tagcgataac gagctcctgc actggatggt ggcgctggat
     4141 ggtaagccgc tggcaagcgg tgaagtgcct ctggatgtcg ctccacaagg taaacagttg
     4201 attgaactgc ctgaactacc gcagccggag agcgccgggc aactctggct cacagtacgc
     4261 gtagtgcaac cgaacgcgac cgcatggtca gaagccgggc acatcagcgc ctggcagcag
     4321 tggcgtctgg cggaaaacct cagtgtgacg ctccccgccg cgtcccacgc catcccgcat
     4381 ctgaccacca gcgaaatgga tttttgcatc gagctgggta ataagcgttg gcaatttaac
     4441 cgccagtcag gctttctttc acagatgtgg attggcgata aaaaacaact gctgacgccg
     4501 ctgcgcgatc agttcacccg tgcaccgctg gataacgaca ttggcgtaag tgaagcgacc
     4561 cgcattgacc ctaacgcctg ggtcgaacgc tggaaggcgg cgggccatta ccaggccgaa
     4621 gcagcgttgt tgcagtgcac ggcagataca cttgctgatg cggtgctgat tacgaccgct
     4681 cacgcgtggc agcatcaggg gaaaacctta tttatcagcc ggaaaaccta ccggattgat
     4741 ggtagtggtc aaatggcgat taccgttgat gttgaagtgg cgagcgatac accgcatccg
     4801 gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa ctggctcgga
     4861 ttagggccgc aagaaaacta tcccgaccgc cttactgccg cctgttttga ccgctgggat
     4921 ctgccattgt cagacatgta taccccgtac gtcttcccga gcgaaaacgg tctgcgctgc
     4981 gggacgcgcg aattgaatta tggcccacac cagtggcgcg gcgacttcca gttcaacatc
     5041 agccgctaca gtcaacagca actgatggaa accagccatc gccatctgct gcacgcggaa
     5101 gaaggcacat ggctgaatat cgacggtttc catatgggga ttggtggcga cgactcctgg
     5161 agcccgtcag tatcggcgga attccagctg agcgccggtc gctaccatta ccagttggtc
     5221 tggtgtcaaa aataagctct tcaggttaga ggtctcgcgg ccgccaccgc tgagcaataa
     5281 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     5341 actatatccg ggtaacgaat tcaagctt
//
