LOCUS       sequence-232205-        5349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Also known as pHH29_32_HG27, one of the pHH29_32 series plasmids. 
            The component plasmid of a unregulated or a regulated device.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5349)
  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 5349)
  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: pHH29_32 series plasmid
FEATURES             Location/Qualifiers
     source          1..5349
                     /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"
     primer_bind     complement(1968..1996)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             3260..3277
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3330..3348)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3344..3391
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4280..4351
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4367..4394
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             4472..4483
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4490..5167
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      5234..5277
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
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 gaacgggcgc gccaaaaaga
     1861 gtattgactt caggaaaatt tttctgcata attatttcaa taacaaactt tgacacttgg
     1921 gtcttatagg atgattaatg cgtaaaggag aagaactgtt cactggagtt gtcccaattc
     1981 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtgagtgga gagggtgaag
     2041 gtgatgcgac atacggcaaa ctgaccctta aatttatttg tactaccggc aagctacctg
     2101 ttccttggcc gacacttgtc actactttcg gttatggtgt tcaatgtttt gcgagatacc
     2161 cagatcatat gaaacagcat gactttttca aaagtgccat gcccgaaggt tatgtacagg
     2221 aaagaactat atttttcaaa gatgacggga actacaaaac acgtgctgaa gttaaatttg
     2281 agggtgatac tctggtaaat agaatcgaat taaaaggtat tgattttaaa gaagatggca
     2341 acattcttgg tcacaagctg gaatacaact acaactcaca caatgtttac atcatggcag
     2401 acaaacaaaa aaatggcatc aaagttaact tcaaaattcg acacaacatt gaagatggaa
     2461 gcgttcaact ggcagaccat tatcaacaga atactccgat tggcgatggc cctgtactct
     2521 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccg aacgaaaaga
     2581 gagatcacat ggtcctgctt gagtttgtaa cagctgctgg gattacacat ggtatggatg
     2641 agctatacaa ataataacaa cctttcacgg agtaggacat caaatgaccg agattcatct
     2701 gcagaccaca gaaagcacca gcgttaatga tggtctgccg ctgaatattg attgggaagg
     2761 tatttttcgt gagcatggcc gtcgtgtcca tcattttatt cgtaaacgtg tcagccatcg
     2821 agaagatgca gaagatctgg aacagatgac ctggctggaa gttcttcgta atagggataa
     2881 atttgccggc gcatcacgtc cggaaacctg ggtttttggt atagcactga acctggtgcg
     2941 taatcatttc cgtctgcaga gcggtcgtcc gcgttgtgat gaactggaag atgatattat
     3001 tctgacccag ggtgatgatc cgagccatat taccgaatat cagcgtctgc tgaatagcac
     3061 cctgaatagt attgcaaacc tgccggaaga tacccgtcgc ctgctgaaca tgctggttga
     3121 aaaagatggt agctatcagg caattgcagc ccatctgaat attccaattg gcaccgttcg
     3181 tagccgtctg agccgtgcac gtgttaccct gaaacagagc gtttttagct aaggatccaa
     3241 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     3301 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     3361 ggcctctaaa cgggtcttga ggggtttttt gtgccttgcg acagacctcc tctctagtat
     3421 ggtgcaaaac ctttcgcggt atggcatgat agcgcctaat acaatgctct aggtagaagc
     3481 aaatgaaaaa cataaatgcc gacgacacat acagaataat taataaaatt aaagcttgta
     3541 gaagcaataa tgatattaat caatgcttat ctgatatgac taaaatggta cattgtgaat
     3601 attatttact cgcgatcatt tatcctcatt ctatggttaa atctgatatt tcaatcctag
     3661 ataattaccc taaaaaatgg aggcaatatt atgatgacgc taatttaata aaatatgatc
     3721 ctatagtaga ttattctaac tccaatcatt caccaattaa ttggaatata tttgaaaaca
     3781 atgctgtaaa taaaaaatct ccaaatgtaa ttaaagaagc gaaaacatca ggtcttatca
     3841 ctgggtttag tttccctatt catacggcta acaatggctt cggaatgctt agttttgcac
     3901 attcagaaaa agacaactat atagatagtt tatttttaca tgcgtgtatg aacataccat
     3961 taattgttcc ttctctagtt gataattatc gaaaaataaa tatagcaaat aataaatcaa
     4021 acaacgattt aaccaaaaga gaaaaagaat gtttagcgtg ggcatgcgaa ggaaaaagct
     4081 cttgggatat ttcaaaaata ttaggttgca gtgagcgtac tgtcactttc catttaacca
     4141 atgcgcaaat gaaactcaat acaacaaacc gctgccaaag tatttctaaa gcaattttaa
     4201 caggagcaat tgattgccca tactttaaaa attaataaca ctgatagtgc tagtgtagat
     4261 cactactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4321 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     4381 tgggcctttc tgcgtttata tactagagac ctgtaggatc gtacaggttt acgcaagaaa
     4441 atggtttgtt atagtcgaat aaatactaga gaaagaggag aaatactaga tggcttcctc
     4501 cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg
     4561 tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc
     4621 taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca
     4681 gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa
     4741 actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgaag acggtggtgt
     4801 tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact
     4861 gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga
     4921 agcttccacc gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg
     4981 tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa
     5041 aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca
     5101 caacgaagac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg
     5161 tgcttaataa cgctgatagt gctagtgtag atcgctacta gagtactagc tctagatgcg
     5221 gccgcgaatt ccagaaatca tccttagcga aagctaagga ttttttttat ctgaaatgcc
     5281 tctgtaacag agcattagcg caaggtgatt tttgtcttct tgcgctaatt ttttggcgtc
     5341 tccactgtg
//
