LOCUS       sequence-440087-        3417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNP561 - T5 Target with T7 Inducible IS621 Recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3417)
  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 3417)
  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: pNP1830
FEATURES             Location/Qualifiers
     source          1..3417
                     /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"
     protein_bind    1570..1588
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1595..1613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1700..1719
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1700..1718
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1719..1743
                     /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             1758..1780
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2773..2790
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2843..2861)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2857..2904
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(2977..3417,1..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 cctttaggga cccctagctg cagcaggtag
     1561 tgctcagtat ctctatcact gatagggatg tcaatctcta tcactgatag ggagaaattg
     1621 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     1681 tggctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
     1741 tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg agcacgaact
     1801 acattatatt ggaatagaca ccgcgaaaga aaagctggat gtggacgtgc tgcgcccaga
     1861 tggtagacac cgtaccaaga agttcgcgaa caccaccaaa ggtcacgatg aacttgtgtc
     1921 ttggctgaag ggccacaaaa tcgatcatgc gcacatctgc attgaagcaa ccggtacata
     1981 catggaaccg gtagcggagt gtctgtacga cgctggttat attgtttccg tgattaaccc
     2041 ggcgttgggc aaggcattcg ctcaaagcga aggtttgcgt aataagaccg ataccgttga
     2101 tgcgcgcatg ctagcggagt tctgccgtca aaaacgccca gcagcttggg aagctccgca
     2161 tccgctggaa cgtgctttgc gcgcactcgt cgtgcgccac caggcgctga cggacatgca
     2221 tacccaagag ctgaatcgta ccgaaaccgc ccgtgaagtt cagcgtccga gcatcgacgc
     2281 ccatctgctg tggctggagg cagagctgaa gcgcctggag aaacaaatta aagacctgac
     2341 tgacgacgac ccggatatga aacatcgtcg caagttgttg gagtcgatcc cgggtattgg
     2401 cgagaaaacg agcgcggttt tgctggcgta catcggctta aaggaccgtt ttgcgcacgc
     2461 gcgccagttt gctgcattcg ctggcctgac tccgcgtcgt tatgaaagcg gctcaagcgt
     2521 tcgtggtgca tctcgtatga gcaaggcggg ccatgttagc ctgcgtcgtg cgttgtacat
     2581 gccggctatg gttgcgacct ccaaaaccga gtggggtcgt gcgtttcgtg atcgtctggc
     2641 ggccaacggt aaaaagggca aggtgatctt aggtgccatg atgcgtaaac tggcccaggt
     2701 tgcatatggc gtcctgaaaa gcggggtccc gtttgatgcg tcccgccaca acccggtggc
     2761 cgcctaatcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
     2821 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2881 aaacgggtct tgaggggttt tttgctgagt tcagttcggc gagcggaaat ggcttacgaa
     2941 cggggcggag gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc
     3001 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     3061 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     3121 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     3181 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     3241 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     3301 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     3361 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacg
//
