LOCUS       sequence-250948-        4423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reporter circuit (PpspA-2G6 with sfgfp::ASV).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4423)
  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 4423)
  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: pLY153
FEATURES             Location/Qualifiers
     source          1..4423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2..59
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(83..100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(362..1312)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1360..1582
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2074..2168)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2198..2989,2990..3058))
                     /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     2821..2840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3059..3163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3228..3246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3250..3293)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3296..3315
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     misc_RNA        3484..3533
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3566..3577
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3584..4297
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    join(4404..4423,1)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
       61 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatctcgag gttacattgt cgatctgttc
      181 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
      241 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
      301 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
      361 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
      421 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
      481 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
      541 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
      601 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
      661 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
      721 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
      781 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
      841 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
      901 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
      961 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     1021 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     1081 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
     1141 tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
     1201 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
     1261 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
     1321 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     1381 atgtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac
     1441 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     1501 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     1561 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     1621 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     1681 ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg
     1741 agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg
     1801 ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa
     1861 gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc
     1921 tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt
     1981 atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac
     2041 aggcttaccc gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg
     2101 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     2161 caggagtcca agcgagctcg taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt cccatggtgc
     3181 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3241 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt
     3301 cgcggccgct tctagaaaag ctgttgtggc aaaaaacgga aagctgttgt ggcaaaaaac
     3361 ggcatcctgg catgttgctg ttgattcttc aatcagatct ttataaatca aaaagataaa
     3421 aaattggcac gcaaattgta ttaacagttc agcaggacaa tcctgaacgc aaatactaga
     3481 gagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac
     3541 aaataatttt gtttaactag agattaaaga ggagaaatac catatgcgta aaggcgaaga
     3601 gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca acggtcataa
     3661 gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt
     3721 catctgtact actggtaaac tgccggtacc ttggccgact ctggtaacga cgctgactta
     3781 tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact tcttcaagtc
     3841 cgccatgccg gaaggctatg tgcaggaacg cacgatttcc tttaaggatg acggcacgta
     3901 caaaacgcgt gcggaagtga aatttgaagg cgataccctg gtaaaccgca ttgagctgaa
     3961 aggcattgac tttaaagaag acggcaatat cctgggccat aagctggaat acaattttaa
     4021 cagccacaat gtttacatca ccgccgataa acaaaaaaat ggcattaaag cgaattttaa
     4081 aattcgccac aacgtggagg atggcagcgt gcagctggct gatcactacc agcaaaacac
     4141 tccaatcggt gatggtcctg ttctgctgcc agacaatcac tatctgagca cgcaaagcgt
     4201 tctgtctaaa gatccgaacg agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc
     4261 agcgggcatc acgcatggta tggatgaact gtacaaaagg cctgctgcaa acgacgaaaa
     4321 ctacgctgca tcagtttaat aatactagag ccaattattg aaggcctccc taacgggggg
     4381 cctttttttg tttctggtct ccctactagt agcggccgct gca
//
