LOCUS       sequence-235335-        2593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Selection of ECF for controller implementation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2593)
  AUTHORS   Huang HH, Qian Y, Del Vecchio D
  TITLE     A quasi-integral controller for adaptation of genetic modules to 
            variable ribosome demand.
  JOURNAL   Nat Commun. 2018 Dec 21;9(1):5415. doi: 10.1038/s41467-018-07899-z.
  PUBMED    30575748
REFERENCE   2  (bases 1 to 2593)
  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 
            Commun."; date: "2018-12-21"; volume: "9(1):5415. doi"; pages: " 
            10.1038/s41467-018-07899-z"
COMMENT     Sequence Label: pHH02_32_1122
FEATURES             Location/Qualifiers
     source          1..2593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(116..118,119..832)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             116..829
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(146..174)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     769..789
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      874..960
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1690..1784)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1815..2474)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2389..2408
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gcgactttat ttaacagcgg catggccagg gaaccgatgc gtcaatcgca ccacacaatg
       61 acaactgctc tcatcattga ggatcctgtc aatttccgcg atagaggagg taaagatgag
      121 caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      181 taatgggcac aaattttctg tccgtggaga gggtgaaggt gatgctacaa acggaaaact
      241 cacccttaaa tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac
      301 tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga
      361 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
      421 tgacgggacc tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
      481 tatcgagtta aagggtattg attttaaaga agatggaaac attcttggac acaaactcga
      541 gtacaacttt aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
      601 agctaacttc aaaattcgcc acaacgttga agatggttcc gttcaactag cagaccatta
      661 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
      721 gacacaatct gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
      781 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat aataaatgtc
      841 cagacctgca ggcatgcaag cctctagagg catcaaataa aacgaaagac tcagtcgaaa
      901 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
      961 ccgccgccct agacctaggg cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1081 caggaaccgt aaaaaggcca ccctcgagga gcgcctcagc gcgccgtagc gtcgataaaa
     1141 attacgggct ggggcgaaac taccatctgt tcgaaaaggt ccgtaaatgg gcctacagag
     1201 cgattcgtca gggctggcct gtattctcac aatggcttga tgccgttatc cagcgtgtcg
     1261 aaatgtacaa cgcttcgctt cccgttccgc tttctccggc tgaatgtcgg gctattggca
     1321 agagcattgc gaaatataca cacaggaaat tctcaccaga gggattttcc gctgtacagg
     1381 ccgctcgcgg tcgcaagggc ggaactaaat ctaagcgcgc agcagttcct acatcagcac
     1441 gttcgctgaa accgtgggag gcattaggca tcagtcgagc gacgtactac cgaaaattaa
     1501 aatgtgaccc agacctcgca aaatgagacc agataagcct tatcagataa cagcgccctt
     1561 ttggcgtctt tttgagcacc taggtgtaaa acacggcagt cttaagctcg ggccctcttt
     1621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgact
     1681 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     1741 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctctcgaa
     1801 ccccagagtc ccgcttacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     1861 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     1921 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     1981 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     2041 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     2101 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     2161 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     2221 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg
     2281 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     2341 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     2401 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     2461 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     2521 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaac
     2581 tcgagtgcca cct
//
