LOCUS       sequence-206253-        7288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose inducible VioB-mCherry expression (stronger RBS). Used to 
            test burden response in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  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 7288)
  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:  pLys-M1
FEATURES             Location/Qualifiers
     source          1..7288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(20..25,26..42,43..48))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      160..704
                     /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     605..624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     821..838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1230..1332
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1333..1992
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1399..1418)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(2308..3186)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3213..3497
                     /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     3420..3439
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(6742..6760)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7010..7029
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 ctagtagata aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaatta
       61 caacttatat cgtatggggc tgacttcagg tgctacattt gaagagataa attgcactga
      121 aatctagaaa tattttatct gattaataag atgatcttct tgagatcgtt ttggtctgcg
      181 cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct
      241 ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact
      301 tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat
      361 taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat
      421 agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct
      481 tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat
      541 aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg
      601 ccaggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
      661 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
      721 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
      781 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
      841 agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct
      901 gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac
      961 tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg
     1021 aacaggaggg acagctgata gaaacagaag ccactggagc acctcaaaaa caccatcata
     1081 cactaaatca gtaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg
     1141 acggaagatc acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc
     1201 tgggccaact tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac
     1261 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     1321 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     1381 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     1441 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     1501 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca
     1561 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     1621 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     1681 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     1741 gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt
     1801 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     1861 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt
     1921 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     1981 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg tgctacgcct
     2041 gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg acccggtcgt
     2101 cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta ccggtttatt
     2161 gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc
     2221 tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc agagcctgat
     2281 aaactctaga agcggccgct tctagagtta tgacaacttg acggctacat cattcacttt
     2341 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     2401 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     2461 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     2521 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     2581 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     2641 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     2701 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     2761 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     2821 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     2881 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     2941 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     3001 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     3061 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     3121 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc
     3181 agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag
     3241 acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct
     3301 tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag
     3361 tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta
     3421 tgccatagca tttttatcca taagattagc ggatcctacc tgacactttt tatcgcaact
     3481 ctctataatt tctccatacc cgtttttttg ggctagcagg catctttcaa ctaaagtaag
     3541 aggtaataat tatgagcatt ctggatttcc cgcgtatcca cttccgtggc tgggcccgtg
     3601 tcaatgcgcc gaccgcgaac cgcgatccgc acggccacat cgatatggcc agcaataccg
     3661 tggcgatggc gggtgagccg ttcgacctgg cacgccatcc tacggagttc caccgtcacc
     3721 tgcgctccct gggtccgcgc ttcggcttgg atggtcgtgc tgacccggaa ggcccgttca
     3781 gcctggccga gggctacaac gctgccggta acaaccactt ttcgtgggag agcgcaaccg
     3841 ttagccacgt gcaatgggat ggcggtgagg cggatcgtgg tgacggtctg gtcggtgctc
     3901 gtttggcact gtggggtcac tacaatgatt atctgcgtac caccttcaat cgtgctcgtt
     3961 gggtcgacag cgacccgacg cgccgtgacg ctgcacaaat ctatgcgggc caattcacca
     4021 ttagcccggc tggtgccggt ccgggtacgc cgtggctgtt tacggcagac attgatgata
     4081 gccatggtgc acgttggacg cgtggcggcc acattgcaga gcgtggcggc cacttcttgg
     4141 atgaagagtt tggtctggca cgcctgtttc agttctctgt gccgaaagat cacccacatt
     4201 ttctgtttca cccgggtccg tttgattccg aggcctggcg tcgtctgcaa ttggctctgg
     4261 aggatgacga cgttctgggt ctgaccgtgc aatatgcgtt gttcaatatg agcaccccgc
     4321 ctcagccgaa cagcccggtt tttcacgata tggtcggtgt tgtcggtctg tggcgtcgtg
     4381 gtgaactggc gagctacccg gctggtcgtc tgctgcgtcc gcgtcaaccg ggtctgggtg
     4441 acctgaccct gcgcgtcaac ggtggtcgcg ttgcgctgaa tttggcgtgt gccattccgt
     4501 tcagcactcg tgccgcgcag ccaagcgcac cggaccgcct gaccccggac ctgggtgcca
     4561 aactgccgct gggcgatctg ctgctgcgtg atgaggacgg cgcactgttg gcacgtgtgc
     4621 cgcaggctct gtaccaagac tattggacga atcacggtat tgtggacctg ccgctgctgc
     4681 gcgaaccgcg tggtagcttg accctgagca gcgaactggc ggagtggcgt gagcaagact
     4741 gggtcaccca aagcgacgcg tctaacctgt acctggaggc accggatcgc cgtcacggtc
     4801 gctttttccc tgagagcatc gcgctgcgca gctactttcg cggtgaagcg cgtgcgcgtc
     4861 cggatatccc gcatcgtatc gagggcatgg gcctggtcgg cgtcgaatct cgtcaggatg
     4921 gcgacgctgc ggaatggcgt ctgacgggtc tgcgtccggg tccggcacgc attgttctgg
     4981 acgatggtgc cgaggcgatc cctctgcgtg ttctgcctga cgattgggcg ctggatgacg
     5041 cgaccgtcga agaagtggat tacgcctttt tgtaccgcca cgttatggcg tattacgagc
     5101 tggtgtatcc attcatgagc gacaaggtgt tttccctggc tgatcgttgc aaatgtgaaa
     5161 cgtacgcacg tctgatgtgg cagatgtgtg atccgcagaa ccgcaacaag tcctattaca
     5221 tgccgagcac ccgcgaactg tcggcaccga aagctcgttt gttcttgaag tatctggccc
     5281 acgtggaagg ccaggcacgc ctgcaagcac ctccgccagc gggtccggca cgcattgaat
     5341 ctaaagccca gttggcggca gagctgcgta aagccgtcga cctggagctg tctgtgatgc
     5401 tgcaatacct gtacgcggcg tatagcattc cgaactatgc acagggccaa caacgtgttc
     5461 gtgacggtgc gtggaccgcc gagcagctgc aactggcgtg cggtagcggt gaccgtcgcc
     5521 gtgatggcgg tattcgtgca gcactgctgg aaattgctca tgaagaaatg attcattacc
     5581 tggtcgttaa caacctgctg atggccctgg gcgagccgtt ctacgcgggt gtcccgctga
     5641 tgggcgaagc ggcacgtcag gcgtttggcc tggacaccga gttcgctctg gaaccgttta
     5701 gcgaaagcac gctggcacgt tttgttcgtc tggaatggcc gcactttatc ccagcaccgg
     5761 gcaaatccat cgcggactgc tatgccgcca ttcgtcaggc gtttttggat ctgccggact
     5821 tgtttggtgg cgaggcaggt aagcgtggcg gtgaacacca cctgttcctg aatgagctga
     5881 ccaaccgtgc gcatccgggt tatcaactgg aagttttcga tcgcgactcg gcgctgtttg
     5941 gtattgcatt tgtgaccgat cagggcgaag gtggcgctct ggacagcccg cactacgaac
     6001 atagccattt tcaacgtctg cgtgaaatga gcgcgcgtat catggctcaa agcgcaccgt
     6061 tcgaaccggc gctgccggcg ttgcgtaatc cggttctgga tgagagcccg ggttgccaac
     6121 gtgtcgcaga cggtcgtgcg cgtgcgctga tggcattgta ccaaggcgtt tatgagctga
     6181 tgtttgcgat gatggcgcag cacttcgccg tgaaaccgct gggtagcttg cgtcgcagcc
     6241 gcctgatgaa cgcagcaatc gatctgatga ccggtctgtt gcgtccgctg agctgcgcgc
     6301 tgatgaacct gccaagcggc atcgccggtc gcacggccgg tccgccgctg ccgggtccgg
     6361 ttgacacccg tagctatgac gactacgcgc tgggctgtcg catgctggca cgccgttgcg
     6421 agcgtctgct ggagcaggcg agcatgctgg aaccgggttg gctgccggat gcgcagatgg
     6481 agctgctgga tttctatcgt cgccaaatgc tggacttggc gtgcggcaaa ctgagccgcg
     6541 aggccgccga agctgcagca aaggaggccg cagctaaaga ggctgcagcg aaagcgatgg
     6601 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     6661 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     6721 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     6781 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     6841 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     6901 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccttg caggacggcg
     6961 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     7021 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     7081 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     7141 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     7201 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     7261 ccgagggccg ccactccacc taataata
//
