LOCUS       sequence-206259-        2844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP cassette under the control of the htpG2 sigma32-responsive 
            promoter. Used to test burden-responsive behaviour of the wt genomic
            promoter. The plasmid has got ColE1 origin of replication and AmpR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2844)
  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 2844)
  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: htpG2-GFP
FEATURES             Location/Qualifiers
     source          1..2844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             100..111
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             118..831
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      843..914
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      930..957
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1081..1872,1873..1941))
                     /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     1704..1723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1942..2046)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2149..2737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2638..2657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccgcctag gccggatcac ctgctctcgc ttgaaattat tctcccttgt ccccatctct
       61 cccacatcct gttttaatta aggatcttag ctactagaga aagaggagaa atactagatg
      121 cgtaaaggcg aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat
      181 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
      241 ctgacgctga agttcatctg tactactggt aaactgccgg ttccttggcc gactctggta
      301 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat
      361 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
      421 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
      481 cgcattgagc tgaaaggcat tgactttaaa gaggacggca atatcctggg ccataagctg
      541 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
      601 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
      661 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
      721 agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg
      781 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa atgaccaggc
      841 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      901 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
      961 ataagtcggt ctcatacagc ggccgcgatt atcaaaaagg atcttcacct agatcctttt
     1021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1081 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1141 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1201 cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa
     1261 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1321 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1381 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1441 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1501 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1561 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1621 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1681 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1741 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1801 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1861 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1921 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1981 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2041 tccgcgcaca tttccccgaa aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt
     2101 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2281 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2761 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2821 ttctgtggat aaccgtgcgg ccgc
//
