LOCUS       sequence-440090-        3448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNP561 - IS621 WT Target with T7 Inducible IS621 Recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3448)
  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 3448)
  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: pNP566
FEATURES             Location/Qualifiers
     source          1..3448
                     /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    1601..1619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1626..1644
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1731..1750
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1731..1749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1750..1774
                     /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             1789..1811
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2804..2821
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2874..2892)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2888..2935
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(3008..3448,1..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 cctgggccgg gtaataccac catcaggcct
     1561 acagatgagc tcgccccaac gaggcaggta gtgctcagta tctctatcac tgatagggat
     1621 gtcaatctct atcactgata gggagaaatt gttatccgct cagagggcac aatcctattc
     1681 cgcgctatcc gacaatctcc aagacattag gtggctcgat cccgcgaaat taatacgact
     1741 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     1801 taagaaggag atataccatg gagcacgaac tacattatat tggaatagac accgcgaaag
     1861 aaaagctgga tgtggacgtg ctgcgcccag atggtagaca ccgtaccaag aagttcgcga
     1921 acaccaccaa aggtcacgat gaacttgtgt cttggctgaa gggccacaaa atcgatcatg
     1981 cgcacatctg cattgaagca accggtacat acatggaacc ggtagcggag tgtctgtacg
     2041 acgctggtta tattgtttcc gtgattaacc cggcgttggg caaggcattc gctcaaagcg
     2101 aaggtttgcg taataagacc gataccgttg atgcgcgcat gctagcggag ttctgccgtc
     2161 aaaaacgccc agcagcttgg gaagctccgc atccgctgga acgtgctttg cgcgcactcg
     2221 tcgtgcgcca ccaggcgctg acggacatgc atacccaaga gctgaatcgt accgaaaccg
     2281 cccgtgaagt tcagcgtccg agcatcgacg cccatctgct gtggctggag gcagagctga
     2341 agcgcctgga gaaacaaatt aaagacctga ctgacgacga cccggatatg aaacatcgtc
     2401 gcaagttgtt ggagtcgatc ccgggtattg gcgagaaaac gagcgcggtt ttgctggcgt
     2461 acatcggctt aaaggaccgt tttgcgcacg cgcgccagtt tgctgcattc gctggcctga
     2521 ctccgcgtcg ttatgaaagc ggctcaagcg ttcgtggtgc atctcgtatg agcaaggcgg
     2581 gccatgttag cctgcgtcgt gcgttgtaca tgccggctat ggttgcgacc tccaaaaccg
     2641 agtggggtcg tgcgtttcgt gatcgtctgg cggccaacgg taaaaagggc aaggtgatct
     2701 taggtgccat gatgcgtaaa ctggcccagg ttgcatatgg cgtcctgaaa agcggggtcc
     2761 cgtttgatgc gtcccgccac aacccggtgg ccgcctaatc gagcaccacc accaccacca
     2821 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     2881 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgag
     2941 ttcagttcgg cgagcggaaa tggcttacga acggggcgga ggggcaccaa taactgcctt
     3001 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     3061 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     3121 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     3181 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     3241 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     3301 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     3361 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     3421 ccagctcacc gtctttcatt gccatacg
//
