LOCUS       sequence-258929-        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"
     promoter        complement(295..329)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      345..372
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        430..548
                     /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             1546..1557
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    1589..1609
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1664..1758
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1879..1898)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1890..2705
                     /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            2872..2980
                     /note="incP origin of transfer"
     CDS             complement(3041..4189)
                     /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      4582..5113
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5240..5326)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5341..5363
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    5388..5409
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(5514..6073,1..277))
                     /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"
ORIGIN
        1 gaaatcgatc cggcgagcga tacagcacat tggtcagaca cagattatcg gtatgttcat
       61 acagatgccg atcatgatcg cgtacgaaac agaccgtgcc accggtgatg gtatagggct
      121 gcccattaaa cacatgaata cccgtgccat gttcgacaat cacaatttca tgaaaatcat
      181 gatgatgttc aggaaaatcc gcctgcggga gccggggttc tatcgccacg gacgcgttac
      241 cggacggaaa aaaatccaca ctatgtaata cggtcatttg ggctccctct agtagctagc
      301 ataataccta ggactgagct agctgtaaac tctagtatca cactggctca ccttcgggtg
      361 ggcctttctg cgtttatata ctagagagac ctttacgccg ctggagcagg aatgcggtga
      421 gcatcacatc accacaattc agcaaattgt gaacatcatc acgttcatct ttccctggtt
      481 gccaatggcc cattttcctg tcagtaacga gaaggtcgcg aattgaggcg ctttttagac
      541 tggtcgtaac tagagtcaca caggaaagta ctagatggca gaatacaaag ataatttact
      601 tggtgaggcg aacagctttc tcgaagtgct ggaacaggtt tcgcatctcg caccgctgga
      661 caaaccggtg ctcatcatcg gcgaacgcgg caccggtaaa gagctgattg ccagccgcct
      721 gcattatctc tcctcccgtt ggcaagggcc gtttatttcc cttaactgcg cggcgttaaa
      781 tgaaaatctg ctggattccg aactgtttgg tcacgaagcg ggggcgttta ccggtgcgca
      841 aaaacgtcat ccagggagat ttgaacgtgc cgacggcggt acgctatttc ttgatgaact
      901 cgctacggca ccgatgatgg tgcaggagaa attattgcgc gtgattgagt acggtgaact
      961 ggagcgcgtt ggcggcagcc aaccattgca ggtgaatgtg cggttggtat gcgcgacgaa
     1021 tgccgatctc ccggcgatgg tcaatgaagg cacttttcgc gctgacctgc tcgaccgact
     1081 ggcttttgat gttgtacaac tgccaccact gcgcgagcgc gaaagcgaca taatgttgat
     1141 ggcagaatac tttgccatcc agatgtgtcg ggaaatcaag ctgcctctgt tcccggggtt
     1201 tacggagcgc gccagagaaa cattgctgaa ttatcgttgg ccgggaaata ttcgtgaatt
     1261 gaaaaacgtg gtggaacgtt cagtgtatcg ccacggcacc agcgattatc cgcttgatga
     1321 catcattatt gatcccttta aacggcgtcc gcctgaagac gctatcgccg tttcagaaac
     1381 cacctcgctt ccaacactgc cgctggattt acgtgagttt cagatgcagc aggaaaaaga
     1441 gttgctgcaa ctcagtttgc aaatgaatgc acgcacacgc cgccgcgaac gtcgcgcaga
     1501 gaaacaggct caatggaaag cagcaaatta atactagagc caattattga aggccgctaa
     1561 cgcggccttt ttttgtttct ggtctcccta ctagtagcgg ccgctgcagg catgcaagct
     1621 tgcggccgcg tcgtgactgg gaaaaccctg gcgactagtc ttggactcct gttgatagat
     1681 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     1741 ttttttattg gtgagaatcc aggggtcccc aataattacg atttaaattt gtgtctcaaa
     1801 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1861 cttacataaa cagtaataca aggggtgtta tgagccatat tcagcgtgaa acgagctgta
     1921 gccgtccgcg tctgaacagc aacatggatg cggatctgta tggctataaa tgggcgcgtg
     1981 ataacgtggg tcagagcggc gcgaccattt atcgtctgta tggcaaaccg gatgcgccgg
     2041 aactgtttct gaaacatggc aaaggcagcg tggcgaacga tgtgaccgat gaaatggtgc
     2101 gtctgaactg gctgaccgaa tttatgccgc tgccgaccat taaacatttt attcgcaccc
     2161 cggatgatgc gtggctgctg accaccgcga ttccgggcaa aaccgcgttt caggtgctgg
     2221 aagaatatcc ggatagcggc gaaaacattg tggatgcgct ggccgtgttt ctgcgtcgtc
     2281 tgcatagcat tccggtgtgc aactgcccgt ttaacagcga tcgtgtgttt cgtctggccc
     2341 aggcgcagag ccgtatgaac aacggcctgg tggatgcgag cgattttgat gatgaacgta
     2401 acggctggcc ggtggaacag gtgtggaaag aaatgcataa actgctgccg tttagcccgg
     2461 atagcgtggt gacccacggc gattttagcc tggataacct gattttcgat gaaggcaaac
     2521 tgattggctg cattgatgtg ggccgtgtgg gcattgcgga tcgttatcag gatctggcca
     2581 ttctgtggaa ctgcctgggc gaatttagcc cgagcctgca aaaacgtctg tttcagaaat
     2641 atggcattga taatccggat atgaacaaac tgcaatttca tctgatgctg gatgaatttt
     2701 tctaataatt aattggaccg cggtccgcgc gttgtccttt tccgctgcat aaccctgctt
     2761 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     2821 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     2881 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     2941 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc gcgatgcagg
     3001 tggctgctga acccccagcc ggaactgacc ccacaaggcc ctagcgtttg caatgcacca
     3061 ggtcatcatt gacccaggcg tgttccacca ggccgctgcc tcgcaactct tcgcaggctt
     3121 cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc cgatccgcac atgaggcgga
     3181 aggtttccag cttgagcggg tacggctccc ggtgcgagct gaaatagtcg aacatccgtc
     3241 gggccgtcgg cgacagcttg cggtacttct cccatatgaa tttcgtgtag tggtcgccag
     3301 caaacagcac gacgatttcc tcgtcgatca ggacctggca acgggacgtt ttcttgccac
     3361 ggtccaggac gcggaagcgg tgcagcagcg acaccgattc caggtgccca acgcggtcgg
     3421 acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat
     3481 accggccatt gatcgaccag cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg
     3541 gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg ctgccacacc agttcgtcat
     3601 cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa
     3661 tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag
     3721 agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca
     3781 aggaaagctg catttccttg atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg
     3841 cttcgctgac ctgttttgcc aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag
     3901 ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac
     3961 gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg gggagccagt tgcacgctgt
     4021 cgcgctcgat cttggccgta gcttgctgga ctatcgagcc gacggactgg aaggtttcgc
     4081 ggggcgcacg catgacggtg cggcttgcga tggtttcggc atcctcggcg gaaaaccccg
     4141 cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt ccgattcatt caccctcctt
     4201 gcgggattgc cccggaatta attccccgga tcgatccgtc gatcttgatc ccctgcgcca
     4261 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     4321 agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag
     4381 ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct
     4441 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     4501 tttctacgtg gctgccattt ttggggtgag gccgttcgcg gccgaggggc gcagcccctg
     4561 gggggatggg aggcccgcgt tagcgggccg ggagggttcg agaagggggg gcacccccct
     4621 tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg tttataaata
     4681 ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct
     4741 tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc
     4801 atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg
     4861 cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg
     4921 ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg
     4981 ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc
     5041 aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg aggtatccac
     5101 aacgccggcg gccctacatg gctctgctgt agtgagtggg ttgcgctccg gcagcggtcc
     5161 tgatcccccg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggcgcgcc
     5221 cagctgtcta gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     5281 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctttaattaa
     5341 agcggataac aatttcacac aggaggccgc ctaggccgcg gccgcgcgaa ttcgcggccg
     5401 cttctagagc ctagttgtct tcatgcatga agacaaaatt aatactagag ggaccaaaac
     5461 gaaaaaagac gcttttcagc gtcttattgt tcgtctttgg taccgagcta gtattattgc
     5521 agaaagccat cccgtccctg gcgaatatca cgcggtgacc agttaaactc tcggcgaaaa
     5581 agcgtcgaaa agtggttact gtcgctgaat ccacagcgat aggcgatgtc agtaacgctg
     5641 gcctcgctgt ggcgtagcag atgtcgggct ttcatcagtc gcaggcggtt caggtatcgc
     5701 tgaggcgtca gtcccgtttg ctgcttaagc tgccgatgta gcgtacgcag tgaaagagaa
     5761 aattgatccg ccacggcatc ccaattcacc tcatcggcaa aatggtcctc cagccaggcc
     5821 agaagcaagt tgagacgtga tgcgctgttt tccaggttct cctgcaaact gcttttacgc
     5881 agcaagagca gtaattgcat aaacaagatc tcgcgactgg cggtcgaggg taaatcattt
     5941 tccccttcct gctgttccat ctgtgcaacc agctgtcgca cctgctgcaa tacgctgtgg
     6001 ttaacgcgcc agtgagacgg atactgccca tccagctctt gtggcagcaa ctgattcagc
     6061 ccggcgagaa act
//
