LOCUS       sequence-243379-        8465 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses protein A and Tn5 Transposase fusion protein in bacterial 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8465)
  AUTHORS   Kaya-Okur HS, Wu SJ, Codomo CA, Pledger ES, Bryson TD, Henikoff JG, 
            Ahmad K, Henikoff S
  TITLE     CUT
  JOURNAL   Nat Commun. 2019 Apr 29;10(1):1930. doi: 10.1038/s41467-019-09982-5.
  PUBMED    31036827
REFERENCE   2  (bases 1 to 8465)
  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"; journalName: "Nat 
            Commun."; date: "2019-04-29"; volume: "10(1):1930. doi"; pages: " 
            10.1038/s41467-019-09982-5"
COMMENT     Sequence Label: 3XFlag-pA-Tn5-Fl
FEATURES             Location/Qualifiers
     source          1..8465
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     297..316
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(336..413)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        join(493..498,499..515,516..521)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        complement(970..1003)
                     /label=T7 Phi-3.8 promoter
                     /note="weak promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1001..1456)
                     /codon_start=1
                     /product="lysozyme from bacteriophage T7"
                     /label=T7 lysozyme
                     /note="inhibitor of T7 RNA polymerase"
                     /translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
                     GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
                     RSLLVTLLAKYEGAVLRAHHEVAPYACPSFDLKRWWEKNELVTSDRG"
     primer_bind     complement(1563..1582)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1896..2555)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2470..2489
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2556..2658)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3585..3804
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3895..4650
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4653..4903
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5229..6404
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6404..7375
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 gaggatggtt aatgatcgtt gcacttgccc ggtgcgaaag cgggatgaga acgggcgata
       61 tagaagaact ttaaaagcgt tgatcaacga aagacgttaa tagggtcgca aactcttaat
      121 gagctcacgg ttgtagagct gcagttagtt gaaacgcagc cgggcgggcc agagatgaac
      181 aggatctgct actttgaaaa gcgttgttgg gcgcgagtag acaggacggg agattgcgga
      241 agtaaatgaa ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
      301 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
      361 tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt gacctagatt
      421 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
      481 taattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt
      541 aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct
      601 cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct
      661 cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct
      721 atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg
      781 ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc
      841 gaccacaccc gtcctgtgga tccggcccat tggctgcctc ccacacttgg atatgcctcc
      901 tcggagcctt atagaattgt ttataagact tgcgcattat ttgacctcca atgcgaacaa
      961 agggaaaccg ctgtggtctc cctttagtga gttcaattaa ttatccacgg tcagaagtga
     1021 ccagttcgtt cttctcccac caacgcttaa ggtcgaacga agggcaagcg tacggcgcca
     1081 cctcatgatg ggcgcgaagc acagcgcctt cgtacttagc cagcagtgtg acaagcagtg
     1141 agcgaaggga ttgcatttgg gctggcgtaa agttagcgtc gaacttacct ttatcgtcga
     1201 taccaccaac aaggcagacg ccgatagagt tgtggttgta acccttagcg tgagagccta
     1261 cagccatctc atctcgtcct gcctccacag taccgtctcg cttgatgata aagtggtatc
     1321 ccacatcgag ccaaccctgc tctttgtgcc actggcgaat ctcacggaca ccaacattct
     1381 gacttggctt ggtagccgag cagtgaacaa agattgcgtc agtagattca cgttgtttaa
     1441 actgtacacg agccattatt tctttcctcc tttccttttt aatctatcaa aggggacccg
     1501 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     1561 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     1621 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     1681 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
     1741 ctgcttccta atgcaggagt cgcataaggg agagcgtcga ccaattctca tgtttgacag
     1801 cttatcatcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca
     1861 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     1921 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     1981 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     2041 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     2101 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     2161 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     2221 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     2281 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
     2341 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     2401 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     2461 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     2521 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     2581 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     2641 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     2701 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
     2761 tcgcgataag ctcatggagc ggcgtaaccg tcgcacagga aggacagaga aagcgcggat
     2821 ctgggaagtg acggacagaa cggtcaggac ctggattggg gaggcggttg ccgccgctgc
     2881 tgctgacggt gtgacgttct ctgttccggt cacaccacat acgttccgcc attcctatgc
     2941 gatgcacatg ctgtatgccg gtataccgct gaaagttctg caaagcctga tgggacataa
     3001 gtccatcagt tcaacggaag tctacacgaa ggtttttgcg ctggatgtgg ctgcccggca
     3061 ccgggtgcag tttgcgatgc cggagtctga tgcggttgcg atgctgaaac aattatcctg
     3121 agaataaatg ccttggcctt tatatggaaa tgtggaactg agtggatatg ctgtttttgt
     3181 ctgttaaaca gagaagctgg ctgttatcca ctgagaagcg aacgaaacag tcgggaaaat
     3241 ctcccattat cgtagagatc cgcattatta atctcaggag cctgtgtagc gtttatagga
     3301 agtagtgttc tgtcatgatg cctgcaagcg gtaacgaaaa cgatttgaat atgccttcag
     3361 gaacaataga aatcttcgtg cggtgttacg ttgaagtgga gcggattatg tcagcaatgg
     3421 acagaacaac ctaatgaaca cagaaccatg atgtggtctg tccttttaca gccagtagtg
     3481 ctcgccgcag tcgagcgaca gggcgaagcc ctcgagtgag cgaggaagca ccagggaaca
     3541 gcacttatat attctgctta cacacgatgc ctgaaaaaac ttcccttggg gttatccact
     3601 tatccacggg gatattttta taattatttt ttttatagtt tttagatctt cttttttaga
     3661 gcgccttgta ggcctttatc catgctggtt ctagagaagg tgttgtgaca aattgccctt
     3721 tcagtgtgac aaatcaccct caaatgacag tcctgtctgt gacaaattgc ccttaaccct
     3781 gtgacaaatt gccctcagaa gaagctgttt tttcacaaag ttatccctgc ttattgactc
     3841 ttttttattt agtgtgacaa tctaaaaact tgtcacactt cacatggatc tgtcatggcg
     3901 gaaacagcgg ttatcaatca caagaaacgt aaaaatagcc cgcgaatcgt ccagtcaaac
     3961 gacctcactg aggcggcata tagtctctcc cgggatcaaa aacgtatgct gtatctgttc
     4021 gttgaccaga tcagaaaatc tgatggcacc ctacaggaac atgacggtat ctgcgagatc
     4081 catgttgcta aatatgctga aatattcgga ttgacctctg cggaagccag taaggatata
     4141 cggcaggcat tgaagagttt cgcggggaag gaagtggttt tttatcgccc tgaagaggat
     4201 gccggcgatg aaaaaggcta tgaatctttt ccttggttta tcaaacgtgc gcacagtcca
     4261 tccagagggc tttacagtgt acatatcaac ccatatctca ttcccttctt tatcgggtta
     4321 cagaaccggt ttacgcagtt tcggcttagt gaaacaaaag aaatcaccaa tccgtatgcc
     4381 atgcgtttat acgaatccct gtgtcagtat cgtaagccgg atggctcagg catcgtctct
     4441 ctgaaaatcg actggatcat agagcgttac cagctgcctc aaagttacca gcgtatgcct
     4501 gacttccgcc gccgcttcct gcaggtctgt gttaatgaga tcaacagcag aactccaatg
     4561 cgcctctcat acattgagaa aaagaaaggc cgccagacga ctcatatcgt attttccttc
     4621 cgcgatatca cttccatgac gacaggatag tctgagggtt atctgtcaca gatttgaggg
     4681 tggttcgtca catttgttct gacctactga gggtaatttg tcacagtttt gctgtttcct
     4741 tcagcctgca tggattttct catacttttt gaactgtaat ttttaaggaa gccaaatttg
     4801 agggcagttt gtcacagttg atttccttct ctttcccttc gtcatgtgac ctgatatcgg
     4861 gggttagttc gtcatcattg atgagggttg attatcacag tttattactc tgaattggct
     4921 atccgcgtgt gtacctctac ctggagtttt tcccacggtg gatatttctt cttgcgctga
     4981 gcgtaagagc tatctgacag aacagttctt ctttgcttcc tcgccagttc gctcgctatg
     5041 ctcggttaca cggctgcggc gagcgctagt gataataagt gactgaggta tgtgctcttc
     5101 ttatctcctt ttgtagtgtt gctcttattt taaacaactt tgcggttttt tgatgacttt
     5161 gcgattttgt tgttgctttg cagtaaattg caagatttaa taaaaaaacg caaagcaatg
     5221 attaaaggat gttcagaatg aaactcatgg aaacacttaa ccagtgcata aacgctggtc
     5281 atgaaatgac gaaggctatc gccattgcac agtttaatga tgacagcccg gaagcgagga
     5341 aaataacccg gcgctggaga ataggtgaag cagcggattt agttggggtt tcttctcagg
     5401 ctatcagaga tgccgagaaa gcagggcgac taccgcaccc ggatatggaa attcgaggac
     5461 gggttgagca acgtgttggt tatacaattg aacaaattaa tcatatgcgt gatgtgtttg
     5521 gtacgcgatt gcgacgtgct gaagacgtat ttccaccggt gatcggggtt gctgcccata
     5581 aaggtggcgt ttacaaaacc tcagtttctg ttcatcttgc tcaggatctg gctctgaagg
     5641 ggctacgtgt tttgctcgtg gaaggtaacg acccccaggg aacagcctca atgtatcacg
     5701 gatgggtacc agatcttcat attcatgcag aagacactct cctgcctttc tatcttgggg
     5761 aaaaggacga tgtcacttat gcaataaagc ccacttgctg gccggggctt gacattattc
     5821 cttcctgtct ggctctgcac cgtattgaaa ctgagttaat gggcaaattt gatgaaggta
     5881 aactgcccac cgatccacac ctgatgctcc gactggccat tgaaactgtt gctcatgact
     5941 atgatgtcat agttattgac agcgcgccta acctgggtat cggcacgatt aatgtcgtat
     6001 gtgctgctga tgtgctgatt gttcccacgc ctgctgagtt gtttgactac acctccgcac
     6061 tgcagttttt cgatatgctt cgtgatctgc tcaagaacgt tgatcttaaa gggttcgagc
     6121 ctgatgtacg tattttgctt accaaataca gcaatagtaa tggctctcag tccccgtgga
     6181 tggaggagca aattcgggat gcctggggaa gcatggttct aaaaaatgtt gtacgtgaaa
     6241 cggatgaagt tggtaaaggt cagatccgga tgagaactgt ttttgaacag gccattgatc
     6301 aacgctcttc aactggtgcc tggagaaatg ctctttctat ttgggaacct gtctgcaatg
     6361 aaattttcga tcgtctgatt aaaccacgct gggagattag ataatgaagc gtgcgcctgt
     6421 tattccaaaa catacgctca atactcaacc ggttgaagat acttcgttat cgacaccagc
     6481 tgccccgatg gtggattcgt taattgcgcg cgtaggagta atggctcgcg gtaatgccat
     6541 tactttgcct gtatgtggtc gggatgtgaa gtttactctt gaagtgctcc ggggtgatag
     6601 tgttgagaag acctctcggg tatggtcagg taatgaacgt gaccaggagc tgcttactga
     6661 ggacgcactg gatgatctca tcccttcttt tctactgact ggtcaacaga caccggcgtt
     6721 cggtcgaaga gtatctggtg tcatagaaat tgccgatggg agtcgccgtc gtaaagctgc
     6781 tgcacttacc gaaagtgatt atcgtgttct ggttggcgag ctggatgatg agcagatggc
     6841 tgcattatcc agattgggta acgattatcg cccaacaagt gcttatgaac gtggtcagcg
     6901 ttatgcaagc cgattgcaga atgaatttgc tggaaatatt tctgcgctgg ctgatgcgga
     6961 aaatatttca cgtaagatta ttacccgctg tatcaacacc gccaaattgc ctaaatcagt
     7021 tgttgctctt ttttctcacc ccggtgaact atctgcccgg tcaggtgatg cacttcaaaa
     7081 agcctttaca gataaagagg aattacttaa gcagcaggca tctaaccttc atgagcagaa
     7141 aaaagctggg gtgatatttg aagctgaaga agttatcact cttttaactt ctgtgcttaa
     7201 aacgtcatct gcatcaagaa ctagtttaag ctcacgacat cagtttgctc ctggagcgac
     7261 agtattgtat aagggcgata aaatggtgct taacctggac aggtctcgtg ttccaactga
     7321 gtgtatagag aaaattgagg ccattcttaa ggaacttgaa aagccagcac cctgatgcga
     7381 ccacgtttta gtctacgttt atctgtcttt acttaatgtc ctttgttaca ggccagaaag
     7441 cataactggc ctgaatattc tctctgggcc cactgttcca cttgtatcgt cggtctgata
     7501 atcagactgg gaccacggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg
     7561 gtcccactcg tatcgtcggt ctgattatta gtctgggacc acggtcccac tcgtatcgtc
     7621 ggtctgataa tcagactggg accacggtcc cactcgtatc gtcggtctga ttattagtct
     7681 gggaccatgg tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact
     7741 cgtatcgtcg gtctgattat tagtctggra ccacggtccc actcgtatcg tcggtctgat
     7801 tattagtctg ggaccacggt cccactcgta tcgtcggtct gattattagt ctgggaccac
     7861 ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacgatccca ctcgtgttgt
     7921 cggtctgatt atcggtctgg gaccacggtc ccacttgtat tgtcgatcag actatcagcg
     7981 tgagactacg attccatcaa tgcctgtcaa gggcaagtat tgacatgtcg tcgtaacctg
     8041 tagaacggag taacctcggt gtgcggttgt atgcctgctg tggattgctg ctgtgtcctg
     8101 cttatccaca acattttgcg cacggttatg tggacaaaat acctggttac ccaggccgtg
     8161 ccggcacgta tttctagtca aacatgagaa ttgtgcctaa tgagtgagct aacttacatt
     8221 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8281 atgaatcggc caacgcgcgg ggagaggcgg tttgcgntat tgggsgccrg ggggggtttt
     8341 cttttnnacc agtgagancg ggssacagyt grttgyscct tcmcccccyk tctcttttas
     8401 wcrtytcgcc arccccmsag wcmrscyssc mtstccgwtt smststtnct cgttgaaaaa
     8461 aaaaa
//
