LOCUS       sequence-257363-        4423 bp    DNA     circular SYN 15-APR-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 Apr 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"
     CDS             complement(255..1205)
                     /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      1253..1475
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(1967..2061)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2091..2882,2883..2951))
                     /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     2714..2733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2952..3056)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3121..3139
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3143..3186)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3189..3208
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     misc_RNA        3377..3426
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3459..3470
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3477..4190
                     /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    4297..4317
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4318..4375
                     /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(4399..4416)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatctc gaggttacat
       61 tgtcgatctg ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg
      121 tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt
      181 aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa
      241 gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt
      301 tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc
      361 aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt
      421 tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca
      481 ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca
      541 acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg
      601 ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac
      661 tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata
      721 tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata
      781 gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac
      841 tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac
      901 ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc
      961 acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc attttcccta
     1021 ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg ttctttcctt
     1081 gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca
     1141 tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca
     1201 gacatacatc tcaattggtc taggtgattt taatcactat accaattgag atgggctagt
     1261 caatgataat tacatgtcct tttcctttga gttgtgggta tctgtaaatt ctgctagacc
     1321 tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag acctttgtgt
     1381 gttttttttg tttatattca agtggttata atttatagaa taaagaaaga ataaaaaaag
     1441 ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt tccgcagtat
     1501 tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta aaaccctaaa
     1561 ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt tgtctccgac
     1621 catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc tcacggctct
     1681 ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg caaggaaact
     1741 acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg cgggtctgct
     1801 atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt ttccagtctg
     1861 accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc agtaaggcag
     1921 cggtatcatc aacaggctta cccgtcttac tgtccctagt gcttggattc tcaccaataa
     1981 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     2041 ctggatctat caacaggagt ccaagcgagc tcgtaaactt ggtctgacag ttaccaatgc
     2101 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2161 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2221 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2281 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2341 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2401 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2461 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2521 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2581 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2641 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2701 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2761 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2821 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2881 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2941 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3001 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3061 tttcccatgg tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3121 aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat
     3181 gatttctgga attcgcggcc gcttctagaa aagctgttgt ggcaaaaaac ggaaagctgt
     3241 tgtggcaaaa aacggcatcc tggcatgttg ctgttgattc ttcaatcaga tctttataaa
     3301 tcaaaaagat aaaaaattgg cacgcaaatt gtattaacag ttcagcagga caatcctgaa
     3361 cgcaaatact agagagctgt caccggatgt gctttccggt ctgatgagtc cgtgaggacg
     3421 aaacagcctc tacaaataat tttgtttaac tagagattaa agaggagaaa taccatatgc
     3481 gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg
     3541 tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac
     3601 tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg actctggtaa
     3661 cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg
     3721 acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg
     3781 atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc
     3841 gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg
     3901 aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta
     3961 aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact
     4021 accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga
     4081 gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg
     4141 agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa aggcctgctg
     4201 caaacgacga aaactacgct gcatcagttt aataatacta gagccaatta ttgaaggcct
     4261 ccctaacggg gggccttttt ttgtttctgg tctccctact agtagcggcc gctgcagtcc
     4321 ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct ttaaaaccga aaagattact
     4381 tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcg
//
