LOCUS       sequence-235306-        3448 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 3448)
  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 3448)
  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: pHH01_32
FEATURES             Location/Qualifiers
     source          1..3448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(106..174,175..966)
                     /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     complement(324..343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      996..1090
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1193..1738
                     /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     1639..1658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1846..1880
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     RBS             1889..1900
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1907..2530
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1980..1999)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2551..2637
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2646..2664
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2671..2689
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2708..2719
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3302..3319
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3372..3390)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3386..3433
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
       61 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      961 tggtaactcg agctgatcct tcaagagctc gcttggactc ctgttgatag atccagtaat
     1021 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     1081 tggtgagaat ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     1141 taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc
     1201 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     1261 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     1321 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     1381 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     1441 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     1501 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     1561 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     1621 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1681 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     1741 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     1801 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgtttat ggctagctca
     1861 gtcctaggta caatgctagc aattcattaa agaggagaaa ggtaccatgt ccagattaga
     1921 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
     1981 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
     2041 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
     2101 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
     2161 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
     2221 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
     2281 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
     2341 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
     2401 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
     2461 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     2521 tgggtcctaa taaacgcgtg ctagaggcat caaataaaac gaaaggctca gtcgaaagac
     2581 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaattac
     2641 tagagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcact
     2701 actagagaaa gaggagaaat actagatgac cgaaattcat ctgcagacca ccgaaagcac
     2761 cagcgttaat gatggtctgc cgctgaatat tgattgggaa ggtatttttc gtgaacatgg
     2821 tcgtcgtgtg catcatttta ttcgtaaacg tgttagccat cgtgaagatg ccgaagatct
     2881 ggaacaaatg acctggctgg aagttctgcg taatcgtgat aaatttgccg gtgcaagccg
     2941 tccggaaacc tgggtttttg gtattgcact gaatctggtg cgtaatcatt ttcgtctgca
     3001 gagcggtcgt ccgcgttgtg atgaactgga agatgatatt attctgaccc agggtgatga
     3061 tccgagccat attaccgaat atcagcgtct gctgaatagc accctgaata gtattgcaaa
     3121 tctgccggaa gatacccgtc gcctgctgaa catgctggtt gaaaaagatg gtagctatca
     3181 ggcaattgca gcccatctga atattccgat tggcaccgtt cgtagccgtc tgagccgtgc
     3241 acgtgttacc ctgaaacaga gcgtttttag ctaaggatcc aagcttgcgg ccgcactcga
     3301 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3361 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3421 gaggggtttt ttgggcgtct ccactgtg
//
