LOCUS       sequence-311607-        3561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PphlF-TA4 in crRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3561)
  AUTHORS   Kuo J, Yuan R, Sanchez C, Paulsson J, Silver PA
  TITLE     Toward a translationally independent RNA-based synthetic oscillator 
            using deactivated CRISPR-Cas.
  JOURNAL   Nucleic Acids Res. 2020 Aug 20;48(14):8165-8177. doi: 
            10.1093/nar/gkaa557.
  PUBMED    32609820
REFERENCE   2  (bases 1 to 3561)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-08-20- 20"; volume: "48"; issue: 
            "14"; pages: "8165-8177"
COMMENT     Sequence Label: pJK517
FEATURES             Location/Qualifiers
     source          1..3561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(302..406)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     455..473
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(477..520)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        994..1028
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     terminator      1768..1839
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1855..1882
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2075..2092)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2209..2754)
                     /direction=LEFT
                     /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     complement(2289..2308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3002..3561,1..232,233..301))
                     /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"
ORIGIN
        1 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
       61 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
      121 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
      181 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
      241 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
      301 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
      361 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
      421 aagtgccacc tgacgtctaa gaaaccattt aattaatagg cgtatcacga ggcagaattt
      481 cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcta aagatctcag
      541 aaatcatttt acaccaacgg gtcacacggg tattatgcta gctaatttct actgttgtag
      601 atcacctagc tcagtcctag gtctcggtac caaattccag aaaagaggcc tcccgaaagg
      661 ggggcctttt ttcgttttgg tccagactgt cgccggatgt gtatccgacc tgacgatggc
      721 ccaaaagggc cgaaacagtc ctctacaaat aattttgttt aagcttttta cacctccaca
      781 actagctggt attatgctag ctaatttcta ctgttgtaga tcaccaacgg gtcacacggg
      841 tctcggtacc aaattccaga aaagagacgc tttcgagcgt cttttttcgt tttggtccgg
      901 atccagtcat aagtctgggc taagcccact gatgagtcgc tgaaatgcga cgaaacttat
      961 gacctctaca aataattttg tttaagatag gattttacgg ctagctcagt cctaggtact
     1021 atgctagcag tacgtctgag cgtgataccc gctcactgaa gatggcccgg tagggccgaa
     1081 acgtacctct acaaataatt ttgtttaact atggactatg tttgaaaggg agaaatacta
     1141 gatggcacgt accccgagcc gtagcagcat tggtagcctg cgtagtccgc atacccataa
     1201 agcaattctg accagcacca ttgaaatcct gaaagaatgt ggttatagcg gtctgagcat
     1261 tgaaagcgtt gcacgtcgtg ccggtgcaag caaaccgacc atttatcgtt ggtggaccaa
     1321 taaagcagca ctgattgccg aagtgtatga aaatgaaagc gaacaggtgc gtaaatttcc
     1381 ggatctgggt agctttaaag ccgatctgga ttttctgctg cgtaatctgt ggaaagtttg
     1441 gcgtgaaacc atttgtggtg aagcatttcg ttgtgttatt gcagaagcac agctggaccc
     1501 tgcaaccctg acccagctga aagatcagtt tatggaacgt cgtcgtgaga tgccgaaaaa
     1561 actggttgaa aatgccatta gcaatggtga actgccgaaa gataccaatc gtgaactgct
     1621 gctggatatg atttttggtt tttgttggta tcgcctgctg accgaacagc tgaccgttga
     1681 acaggatatt gaagaattta ccttcctgct gattaatggt gtttgtccgg gtacacagcg
     1741 ttaactaggg cccatacccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1801 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     1861 ccttcgggtg ggcctttctg cgtttatata cgcgacgtac ggtggaatct gattcgttac
     1921 caattgacat gatacgaaac gtaccgtatc gttaaggtta atttctactg ttgtagatca
     1981 cctccacaac tagctggtct cggtaccaaa gacgaacaat aagacgctga aaagcgtctt
     2041 ttttcgtttt ggtccttaat taactgcagg cttcctcgct cactgactcg ctacgctcgg
     2101 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     2161 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     2221 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     2281 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     2341 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     2401 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     2461 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     2521 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     2581 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     2641 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     2701 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     2761 catcttatta atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta
     2821 gcacctgaag tcagccccat acgatataag ttgttactag tgcttggcag tggaacgaaa
     2881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3541 ctgtgactgg tgagtactca a
//
