LOCUS       sequence-210731-        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             91..102
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             109..822
                     /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      834..905
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      921..948
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1072..1863,1864..1932))
                     /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     1695..1714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1933..2037)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2140..2728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2629..2648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccggatca cctgctctcg cttgaaatta ttctcccttg tccccatctc tcccacatcc
       61 tgttttaatt aaggatctta gctactagag aaagaggaga aatactagat gcgtaaaggc
      121 gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga tgtcaacggt
      181 cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg
      241 aagttcatct gtactactgg taaactgccg gttccttggc cgactctggt aacgacgctg
      301 acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca tgacttcttc
      361 aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa ggatgacggc
      421 acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa ccgcattgag
      481 ctgaaaggca ttgactttaa agaggacggc aatatcctgg gccataagct ggaatacaat
      541 tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat taaagcgaat
      601 tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca ctaccagcaa
      661 aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct gagcacgcaa
      721 agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct ggagttcgta
      781 accgcagcgg gcatcacgca tggtatggat gaactgtaca aatgaccagg catcaaataa
      841 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
      901 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tataagtcgg
      961 tctcatacag cggccgcgat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1021 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1081 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1141 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1201 aatgataccg cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1261 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1321 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1381 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1441 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1501 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1561 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1621 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1681 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1741 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1801 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1861 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1921 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1981 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2041 atttccccga aaagtgccac ctgtcatgac caaaatccct taacgtgagt tttcgttcca
     2101 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2161 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2221 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2281 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2341 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2401 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2461 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2521 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2581 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2641 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2701 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2761 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2821 taaccgtgcg gccgcggccg ccta
//
