LOCUS       sequence-440086-        3869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNP1950 - J23119-IS621 bRNA1 TBL-lacZ-1 (Extended RTG), DBL-WT, HDV;
            T7-IS621 Recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3869)
  AUTHORS   Durrant MG, Perry NT, Pai JJ, Jangid AR, Athukoralage JS, Hiraizumi 
            M, McSpedon JP, Pawluk A, Nishimasu H, Konermann S, Hsu PD
  TITLE     Bridge RNAs direct programmable recombination of target and donor 
            DNA.
  JOURNAL   Nature. 2024 Jun;630(8018):984-993. doi: 10.1038/s41586-024-07552-4.
            Epub 2024 Jun 26.
  PUBMED    38926615
REFERENCE   2  (bases 1 to 3869)
  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: 
            "10.1038/s41586-024-07552-4"; journalName: "Nature"; date: 
            "2024-06"; volume: "630"; issue: "8018"; pages: "984-993"
COMMENT     Sequence Label: pNP2270
FEATURES             Location/Qualifiers
     source          1..3869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /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(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1534..1568
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     ncRNA           1746..1813
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     primer_bind     2152..2171
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2152..2170
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2171..2195
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             2210..2232
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3225..3242
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3295..3313)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3309..3356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(3429..3869,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagcttg cctttgacag ctagctcagt cctaggtata
     1561 atgctagcag tgcagagaaa atcggccagt tttctctgcc tgcagtccgc atgccgtgaa
     1621 gtgccttggg ttctaacctg ttccagagat ttatgcagcg gactgccttt ctcccaaagt
     1681 gataaaccgg acagtatcat ggaccggttt tcccggtaat ccgtatttgc aaggttggtt
     1741 tcactggccg gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca tgcttcggca
     1801 tggcgaatgg gacgatctga cagctgtcag cccgcctaat gagcgggctt ttttttcatg
     1861 agaattacaa cttataattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     1921 ccccatacga tataagttgt ttcagccaaa aaacttaaga ccgccggtct tgtccactac
     1981 cttgcagtaa tgcggtggac aggatcggcg gttttctttt ctcttctcaa ccctgaattc
     2041 gcatctagac tagcgcttga atttcgaaat tgttatccgc tcagagggca caatcctatt
     2101 ccgcgctatc cgacaatctc caagacatta ggtggctcga tcccgcgaaa ttaatacgac
     2161 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
     2221 ttaagaagga gatataccat ggagcacgaa ctacattata ttggaataga caccgcgaaa
     2281 gaaaagctgg atgtggacgt gctgcgccca gatggtagac accgtaccaa gaagttcgcg
     2341 aacaccacca aaggtcacga tgaacttgtg tcttggctga agggccacaa aatcgatcat
     2401 gcgcacatct gcattgaagc aaccggtaca tacatggaac cggtagcgga gtgtctgtac
     2461 gacgctggtt atattgtttc cgtgattaac ccggcgttgg gcaaggcatt cgctcaaagc
     2521 gaaggtttgc gtaataagac cgataccgtt gatgcgcgca tgctagcgga gttctgccgt
     2581 caaaaacgcc cagcagcttg ggaagctccg catccgctgg aacgtgcttt gcgcgcactc
     2641 gtcgtgcgcc accaggcgct gacggacatg catacccaag agctgaatcg taccgaaacc
     2701 gcccgtgaag ttcagcgtcc gagcatcgac gcccatctgc tgtggctgga ggcagagctg
     2761 aagcgcctgg agaaacaaat taaagacctg actgacgacg acccggatat gaaacatcgt
     2821 cgcaagttgt tggagtcgat cccgggtatt ggcgagaaaa cgagcgcggt tttgctggcg
     2881 tacatcggct taaaggaccg ttttgcgcac gcgcgccagt ttgctgcatt cgctggcctg
     2941 actccgcgtc gttatgaaag cggctcaagc gttcgtggtg catctcgtat gagcaaggcg
     3001 ggccatgtta gcctgcgtcg tgcgttgtac atgccggcta tggttgcgac ctccaaaacc
     3061 gagtggggtc gtgcgtttcg tgatcgtctg gcggccaacg gtaaaaaggg caaggtgatc
     3121 ttaggtgcca tgatgcgtaa actggcccag gttgcatatg gcgtcctgaa aagcggggtc
     3181 ccgtttgatg cgtcccgcca caacccggtg gccgcctaat cgagcaccac caccaccacc
     3241 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     3301 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     3361 gttcagttcg gcgagcggaa atggcttacg aacggggcgg aggggcacca ataactgcct
     3421 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     3481 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     3541 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     3601 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     3661 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     3721 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     3781 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     3841 accagctcac cgtctttcat tgccatacg
//
