LOCUS       sequence-250933-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pspF-Delta-HTH::lambda-N22plus gene controlled by promoter PrhaB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  AUTHORS   Liu Y, Wan X, Wang B
  TITLE     Engineered CRISPRa enables programmable eukaryote-like gene 
            activation in bacteria.
  JOURNAL   Nat Commun. 2019 Aug 26;10(1):3693. doi: 10.1038/s41467-019-11479-0.
  PUBMED    31451697
REFERENCE   2  (bases 1 to 6073)
  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: "2019-08-26"; volume: "10(1):3693. doi"; pages: " 
            10.1038/s41467-019-11479-0"
COMMENT     Sequence Label: pLY151
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    56..77
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(182..1018)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        complement(1036..1070)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      1086..1113
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1171..1289
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             2287..2298
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2330..2350
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2405..2499
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2620..2639)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2631..3446
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            3613..3721
                     /note="incP origin of transfer"
     CDS             complement(3782..4930)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      5323..5854
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5981..6067)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgcggccgct tctagagcct agttgtcttc atgcatgaag acaaaattaa tactagaggg
      121 accaaaacga aaaaagacgc ttttcagcgt cttattgttc gtctttggta ccgagctagt
      181 attattgcag aaagccatcc cgtccctggc gaatatcacg cggtgaccag ttaaactctc
      241 ggcgaaaaag cgtcgaaaag tggttactgt cgctgaatcc acagcgatag gcgatgtcag
      301 taacgctggc ctcgctgtgg cgtagcagat gtcgggcttt catcagtcgc aggcggttca
      361 ggtatcgctg aggcgtcagt cccgtttgct gcttaagctg ccgatgtagc gtacgcagtg
      421 aaagagaaaa ttgatccgcc acggcatccc aattcacctc atcggcaaaa tggtcctcca
      481 gccaggccag aagcaagttg agacgtgatg cgctgttttc caggttctcc tgcaaactgc
      541 ttttacgcag caagagcagt aattgcataa acaagatctc gcgactggcg gtcgagggta
      601 aatcattttc cccttcctgc tgttccatct gtgcaaccag ctgtcgcacc tgctgcaata
      661 cgctgtggtt aacgcgccag tgagacggat actgcccatc cagctcttgt ggcagcaact
      721 gattcagccc ggcgagaaac tgaaatcgat ccggcgagcg atacagcaca ttggtcagac
      781 acagattatc ggtatgttca tacagatgcc gatcatgatc gcgtacgaaa cagaccgtgc
      841 caccggtgat ggtatagggc tgcccattaa acacatgaat acccgtgcca tgttcgacaa
      901 tcacaatttc atgaaaatca tgatgatgtt caggaaaatc cgcctgcggg agccggggtt
      961 ctatcgccac ggacgcgtta ccggacggaa aaaaatccac actatgtaat acggtcattt
     1021 gggctccctc tagtagctag cataatacct aggactgagc tagctgtaaa ctctagtatc
     1081 acactggctc accttcgggt gggcctttct gcgtttatat actagagaga cctttacgcc
     1141 gctggagcag gaatgcggtg agcatcacat caccacaatt cagcaaattg tgaacatcat
     1201 cacgttcatc tttccctggt tgccaatggc ccattttcct gtcagtaacg agaaggtcgc
     1261 gaattgaggc gctttttaga ctggtcgtaa ctagagtcac acaggaaagt actagatggc
     1321 agaatacaaa gataatttac ttggtgaggc gaacagcttt ctcgaagtgc tggaacaggt
     1381 ttcgcatctc gcaccgctgg acaaaccggt gctcatcatc ggcgaacgcg gcaccggtaa
     1441 agagctgatt gccagccgcc tgcattatct ctcctcccgt tggcaagggc cgtttatttc
     1501 ccttaactgc gcggcgttaa atgaaaatct gctggattcc gaactgtttg gtcacgaagc
     1561 gggggcgttt accggtgcgc aaaaacgtca tccagggaga tttgaacgtg ccgacggcgg
     1621 tacgctattt cttgatgaac tcgctacggc accgatgatg gtgcaggaga aattattgcg
     1681 cgtgattgag tacggtgaac tggagcgcgt tggcggcagc caaccattgc aggtgaatgt
     1741 gcggttggta tgcgcgacga atgccgatct cccggcgatg gtcaatgaag gcacttttcg
     1801 cgctgacctg ctcgaccgac tggcttttga tgttgtacaa ctgccaccac tgcgcgagcg
     1861 cgaaagcgac ataatgttga tggcagaata ctttgccatc cagatgtgtc gggaaatcaa
     1921 gctgcctctg ttcccggggt ttacggagcg cgccagagaa acattgctga attatcgttg
     1981 gccgggaaat attcgtgaat tgaaaaacgt ggtggaacgt tcagtgtatc gccacggcac
     2041 cagcgattat ccgcttgatg acatcattat tgatcccttt aaacggcgtc cgcctgaaga
     2101 cgctatcgcc gtttcagaaa ccacctcgct tccaacactg ccgctggatt tacgtgagtt
     2161 tcagatgcag caggaaaaag agttgctgca actcagtttg caaatgaatg cacgcacacg
     2221 ccgccgcgaa cgtcgcgcag agaaacaggc tcaatggaaa gcagcaaatt aatactagag
     2281 ccaattattg aaggccgcta acgcggcctt tttttgtttc tggtctccct actagtagcg
     2341 gccgctgcag gcatgcaagc ttgcggccgc gtcgtgactg ggaaaaccct ggcgactagt
     2401 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     2461 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc caataattac
     2521 gatttaaatt tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     2581 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     2641 ttcagcgtga aacgagctgt agccgtccgc gtctgaacag caacatggat gcggatctgt
     2701 atggctataa atgggcgcgt gataacgtgg gtcagagcgg cgcgaccatt tatcgtctgt
     2761 atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg caaaggcagc gtggcgaacg
     2821 atgtgaccga tgaaatggtg cgtctgaact ggctgaccga atttatgccg ctgccgacca
     2881 ttaaacattt tattcgcacc ccggatgatg cgtggctgct gaccaccgcg attccgggca
     2941 aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg cgaaaacatt gtggatgcgc
     3001 tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg caactgcccg tttaacagcg
     3061 atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa caacggcctg gtggatgcga
     3121 gcgattttga tgatgaacgt aacggctggc cggtggaaca ggtgtggaaa gaaatgcata
     3181 aactgctgcc gtttagcccg gatagcgtgg tgacccacgg cgattttagc ctggataacc
     3241 tgattttcga tgaaggcaaa ctgattggct gcattgatgt gggccgtgtg ggcattgcgg
     3301 atcgttatca ggatctggcc attctgtgga actgcctggg cgaatttagc ccgagcctgc
     3361 aaaaacgtct gtttcagaaa tatggcattg ataatccgga tatgaacaaa ctgcaatttc
     3421 atctgatgct ggatgaattt ttctaataat taattggacc gcggtccgcg cgttgtcctt
     3481 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     3541 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     3601 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     3661 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     3721 gtaggccggc cgcgatgcag gtggctgctg aacccccagc cggaactgac cccacaaggc
     3781 cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc
     3841 ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat
     3901 ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc
     3961 tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga
     4021 atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc
     4081 aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt
     4141 ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc
     4201 attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct
     4261 cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct
     4321 gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca
     4381 cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt
     4441 tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg
     4501 gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt
     4561 tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc caggtcctcg ccggcggttt
     4621 ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg
     4681 ccgcctcctg ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag
     4741 ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg actatcgagc
     4801 cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg
     4861 catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg
     4921 tccgattcat tcaccctcct tgcgggattg ccccggaatt aattccccgg atcgatccgt
     4981 cgatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
     5041 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt
     5101 ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct
     5161 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     5221 gcaccgtttc tgcggactgg ctttctacgt ggctgccatt tttggggtga ggccgttcgc
     5281 ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc
     5341 gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt
     5401 taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc
     5461 cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa
     5521 atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg
     5581 cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca
     5641 ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg
     5701 aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg
     5761 cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca
     5821 agttttccgc gaggtatcca caacgccggc ggccctacat ggctctgctg tagtgagtgg
     5881 gttgcgctcc ggcagcggtc ctgatccccc gcagaaaaaa aggatctcaa gaagatcctt
     5941 tgatcttttc tacggcgcgc ccagctgtct agggcggcgg atttgtccta ctcaggagag
     6001 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     6061 ttatttgatg cct
//
