LOCUS       sequence-440088-        3715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23119-BsaI_bRNA Golden Gate Landing Pad-HDV; T7-IS621 Recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3715)
  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 3715)
  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: pNP1950
FEATURES             Location/Qualifiers
     source          1..3715
                     /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           1592..1659
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     primer_bind     1998..2017
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1998..2016
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2017..2041
                     /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             2056..2078
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3071..3088
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3141..3159)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3155..3202
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(3275..3715,1..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 atgctagccg agaccatggc caccggtctc aggccggcat ggtcccagcc tcctcgctgg
     1621 cgccggctgg gcaacatgct tcggcatggc gaatgggacg atctgacagc tgtcagcccg
     1681 cctaatgagc gggctttttt ttcatgagaa ttacaactta taatttcagt gcaatttatc
     1741 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtttca gccaaaaaac
     1801 ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg gtggacagga tcggcggttt
     1861 tcttttctct tctcaaccct gaattcgcat ctagactagc gcttgaattt cgaaattgtt
     1921 atccgctcag agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg
     1981 gctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     2041 ccctctagaa ataattttgt ttaactttaa gaaggagata taccatggag cacgaactac
     2101 attatattgg aatagacacc gcgaaagaaa agctggatgt ggacgtgctg cgcccagatg
     2161 gtagacaccg taccaagaag ttcgcgaaca ccaccaaagg tcacgatgaa cttgtgtctt
     2221 ggctgaaggg ccacaaaatc gatcatgcgc acatctgcat tgaagcaacc ggtacataca
     2281 tggaaccggt agcggagtgt ctgtacgacg ctggttatat tgtttccgtg attaacccgg
     2341 cgttgggcaa ggcattcgct caaagcgaag gtttgcgtaa taagaccgat accgttgatg
     2401 cgcgcatgct agcggagttc tgccgtcaaa aacgcccagc agcttgggaa gctccgcatc
     2461 cgctggaacg tgctttgcgc gcactcgtcg tgcgccacca ggcgctgacg gacatgcata
     2521 cccaagagct gaatcgtacc gaaaccgccc gtgaagttca gcgtccgagc atcgacgccc
     2581 atctgctgtg gctggaggca gagctgaagc gcctggagaa acaaattaaa gacctgactg
     2641 acgacgaccc ggatatgaaa catcgtcgca agttgttgga gtcgatcccg ggtattggcg
     2701 agaaaacgag cgcggttttg ctggcgtaca tcggcttaaa ggaccgtttt gcgcacgcgc
     2761 gccagtttgc tgcattcgct ggcctgactc cgcgtcgtta tgaaagcggc tcaagcgttc
     2821 gtggtgcatc tcgtatgagc aaggcgggcc atgttagcct gcgtcgtgcg ttgtacatgc
     2881 cggctatggt tgcgacctcc aaaaccgagt ggggtcgtgc gtttcgtgat cgtctggcgg
     2941 ccaacggtaa aaagggcaag gtgatcttag gtgccatgat gcgtaaactg gcccaggttg
     3001 catatggcgt cctgaaaagc ggggtcccgt ttgatgcgtc ccgccacaac ccggtggccg
     3061 cctaatcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     3121 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     3181 acgggtcttg aggggttttt tgctgagttc agttcggcga gcggaaatgg cttacgaacg
     3241 gggcggaggg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     3301 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     3361 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     3421 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     3481 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     3541 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     3601 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     3661 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacg
//
