LOCUS       sequence-468573-        8025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOBK360_VM_N3 assembled plasmid (Kanamycin R) containing two 
            inserts (POC1542-POC1543) assembled in POC1519 using the 
            site-selective methylation protection approach.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8025)
  AUTHORS   Flores-Fernandez CN, Lin D, Robins K, O'Callaghan CA
  TITLE     UniClo: scarless hierarchical DNA assembly without sequence 
            constraint.
  JOURNAL   Nucleic Acids Res. 2025 Jun 20;53(12):gkaf548. doi: 
            10.1093/nar/gkaf548.
  PUBMED    40548934
REFERENCE   2  (bases 1 to 8025)
  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: "Nucleic 
            Acids Res."; date: "2025-06-20"; volume: "53(12):gkaf548. doi"; 
            pages: " 10.1093/nar/gkaf548"
COMMENT     Sequence Label: POC1548
FEATURES             Location/Qualifiers
     source          1..8025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2000..2104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2105..2899
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3714..3933
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             4024..4779
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4782..5032
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5358..6533
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6533..7504
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccacgg tcccactcgt
       61 atcgtcggtc tgattattag tctgggacca cgatcccact cgtgttgtcg gtctgattat
      121 cggtctggga ccacggtccc acttgtattg tcgatcagac tatcagcgtg agactacgat
      181 tccatcaatg cctgtcaagg gcaagtattg acatgtcgtc gtaacctgta gaacggagta
      241 acctcggtgt gcggttgtat gcctgctgtg gattgctgct gtgtcctgct tatccacaac
      301 attttgcgca cggttatgtg gacaaaatac ctggttaccc aggccgtgcc ggcacgttaa
      361 ccgggctgca tccgatgcaa gtgtgtcgct gtcgacgcaa gcggccgcac actctgtggt
      421 gcagtacctc tcacgacttg gtctccgaaa tgcggctcca atcttcctcc tgaggggtct
      481 gggcctttgg aaatcaaacg ctgtcagact gggttgctgg acgattctgt tcacatttac
      541 aatgggacaa gggaaacaag gaggccccca agtgaatctc tgggttccac acaaactcct
      601 cctgccctta ctgtgtatca gcagccctgc ctcgtcctgg ggatgagggt ccatcactcc
      661 tgcctggaga ggaggggagt gctcttcttc cctgcttgtc tctaggccca tacagtcctg
      721 tgggcaactg taatgtgtag ctcaatgggc tcttgtttgt ccccttgtct gagtgcttcc
      781 atcccttagt gatgtctctg attcaataaa tgcaccgtgc cctgaaattt gtgtgaaaag
      841 ggggatgaga cagggtaaga gagagacatt ttaccttttg aacattttga attttgaatt
      901 ttgaactata tcaagaaata aaagataatt cctaaggaga ccaaacaaac ccccaaaaaa
      961 ttcaaaatga aaaacttttt aaaaactaat ggaatttttt aacctttatc gaaataaacc
     1021 ttgagggcaa ggagggttta tttcctctta tctcagtccc agctcctctt aaaacaatgc
     1081 cctgcacaca gtaggtattt gataaatgtt taccaaatga agggattgcc tggaatggct
     1141 tggcagacag gaaagcagaa tgagctgtac tctcagctac tcgaagctgg tccctgcttt
     1201 aggctgcgct ccctttctgg gccccaggtc ccgccttcta aatctcccca ggtctccagc
     1261 ccactggaat tttctcttcc aagcgtggcc ccgccctctc cgctcgtgat tggccctaag
     1321 ttccgggccc cagtttcatt ggtcctccgc cacacttcag ccctgcttcc ccgtcacttt
     1381 tcagtcctcc tcgggatcgc gcatcacctg cactttctgg tctcctcctg ctctttctct
     1441 cctcgcgtct cctccgcttc ctctcacttt tcggacaaac cagtccttct gaggcccatg
     1501 ggttcccggg ctgcctccgg ggctgctcct gtgaatggca ttcgagtgcc cttgttcgag
     1561 ctccttatat gtttcagtta ctgatccttt gtcagatgaa tagtttgaaa atattttctc
     1621 ccattcttgg atggtctctt cattttgttt attgtttcct ttgctgtgca gaagcctttt
     1681 tacttgatat gatcccattt atgcaatttt actttggtta cctcccttgc ccctgcacac
     1741 ccagtccttg cacctggcct acaagtttgc catcttcatt cccccttctt ctgttcatca
     1801 gccccctcct ctatcctccc accctcacag ttttctttgt atatgaaatc ctcgttcttg
     1861 tccctttgcc cgtgtgcatt tcctgcccca ggaaggttgg gacagcagac ctgtgtgtta
     1921 aacatcaatg tgaagttact tccaggtgag accaagtcgt gagaggtact gcaccacgaa
     1981 gtgtgcggcc gccttcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2101 gagtatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg
     2161 cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga
     2221 aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg
     2281 ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt
     2341 tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc
     2401 ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat
     2461 caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg
     2521 cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg
     2581 ggaagaggac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa
     2641 gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa
     2701 agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta
     2761 tgacattgcc ttctgcgtcc ggtcgctcag ggaggatatc ggggaagaac agtatgtcga
     2821 gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt
     2881 actggatgaa ttgttttagg agctggagcg gcgtaaccgt cgcacaggaa ggacagagaa
     2941 agcgcggatc tgggaagtga cggacagaac ggtcaggacc tggattgggg aggcggttgc
     3001 cgccgctgct gctgacggtg tgacgttctc tgttccggtc acaccacata cgttccgcca
     3061 ttcctatgcg atgcacatgc tgtatgccgg tataccgctg aaagttctgc aaagcctgat
     3121 gggacataag tccatcagtt caacggaagt ctacacgaag gtttttgcgc tggatgtggc
     3181 tgcccggctc cgggtgcagt ttgcgatgcc ggagtctgat gcggttgcga tgctgaaaca
     3241 attatcctga gaataaatgc cttggccttt atatggaaat gtggaactga gtggatatgc
     3301 tgtttttgtc tgttaaacag agaagctggc tgttatccac tgagaagcga acgaaacagt
     3361 cgggaaaatc tcccattatc gtagagatcc gcattattaa tctcaggagc ctgtgtagcg
     3421 tttataggaa gtagtgttct gtcatgatgc ctgcaagcgg taacgaaaac gatttgaata
     3481 tgccttcagg aacaatagaa atcttcgtgc ggtgttacgt tgaagtggag cggattatgt
     3541 cagcaatgga cagaacaacc taatgaacac agaaccatga tgtggtctgt ccttttacag
     3601 ccagtagtgc tcgccgcagt cgagcgacag ggcgaagccc tcgagtgagc gaggaagcac
     3661 cagggaacag cacttatata ttctgcttac acacgatgcc tgaaaaaact tcccttgggg
     3721 ttatccactt atccacgggg atatttttat aattattttt tttatagttt ttagatcttc
     3781 ttttttagag cgccttgtag gcctttatcc atgctggttc tagagaaggt gttgtgacaa
     3841 attgcccttt cagtgtgaca aatcaccctc aaatgacagt cctgtctgtg acaaattgcc
     3901 cttaaccctg tgacaaattg ccctcagaag aagctgtttt ttcacaaagt tatccctgct
     3961 tattgactct tttttattta gtgtgacaat ctaaaaactt gtcacacttc acatggatct
     4021 gtcatggcgg aaacagcggt tatcaatcac aagaaacgta aaaatagccc gcgaatcgtc
     4081 cagtcaaacg acctcactga ggcggcatat agtctctccc gggatcaaaa acgtatgctg
     4141 tatctgttcg ttgaccagat cagaaaatct gatggcaccc tacaggaaca tgacggtatc
     4201 tgcgagatcc atgttgctaa atatgctgaa atattcggat tgacctctgc ggaagccagt
     4261 aaggatatac ggcaggcatt gaagagtttc gcggggaagg aagtggtttt ttatcgccct
     4321 gaagaggatg ccggcgatga aaaaggctat gaatcttttc cttggtttat caaacgtgcg
     4381 cacagtccat ccagagggct ttacagtgta catatcaacc catatctcat tcccttcttt
     4441 atcgggttac agaaccggtt tacgcagttt cggcttagtg aaacaaaaga aatcaccaat
     4501 ccgtatgcca tgcgtttata cgaatccctg tgtcagtatc gtaagccgga tggctcaggc
     4561 atcgtgtctc tgaaaatcga ctggatcata gagcgttacc agctgcctca aagttaccag
     4621 cgtatgcctg acttccgccg ccgcttcctg caggtctgtg ttaatgagat caacagcaga
     4681 actccaatgc gcctctcata cattgagaaa aagaaaggcc gccagacgac tcatatcgta
     4741 ttttccttcc gcgatatcac ttccatgacg acaggatagt ctgagggtta tctgtcacag
     4801 atttgagggt ggttcgtcac atttgttctg acctactgag ggtaatttgt cacagttttg
     4861 ctgtttcctt cagcctgcat ggattttctc atactttttg aactgtaatt tttaaggaag
     4921 ccaaatttga gggcagtttg tcacagttga tttccttctc tttcccttcg tcatgtgacc
     4981 tgatatcggg ggttagttcg tcatcattga tgagggttga ttatcacagt ttattactct
     5041 gaattggcta tccgcgtgtg tacctctacc tggagttttt cccacggtgg atatttcttc
     5101 ttgcgctgag cgtaagagct atctgacaga acagttcttc tttgcttcct cgccagttcg
     5161 ctcgctatgc tcggttacac ggctgcggcg agcgctagtg ataataagtg actgaggtat
     5221 gtggtgttct tatctccttt tgtagtgttg ctcttatttt aaacaacttt gcggtttttt
     5281 gatgactttg cgattttgtt gttgctttgc agtaaattgc aagatttaat aaaaaaacgc
     5341 aaagcaatga ttaaaggatg ttcagaatga aactcatgga aacacttaac cagtgcataa
     5401 acgctggtca tgaaatgacg aaggctatcg ccattgcaca gtttaatgat gacagcccgg
     5461 aagcgaggaa aataacccgg cgctggagaa taggtgaagc agcggattta gttggggttt
     5521 cttctcaggc tatcagagat gccgagaaag cagggcgact accgcacccg gatatggaaa
     5581 ttcgaggacg ggttgagcaa cgtgttggtt atacaattga acaaattaat catatgcgtg
     5641 atgtgtttgg tacgcgattg cgacgtgctg aggacgtatt tccaccggtg atcggggttg
     5701 ctgcccataa aggtggcgtt tacaaaacct cagtttctgt tcatcttgct caggatctgg
     5761 ctctgaaggg gctacgtgtt ttgctcgtgg aaggtaacga cccccaggga acagcctcaa
     5821 tgtatcacgg atgggtacca gatcttcata ttcatgcaga agatactctc ctgcctttct
     5881 atcttgggga aaaggacgat gtcacttatg caataaagcc cacttgctgg ccggggcttg
     5941 acattattcc ttcctgtctg gctctgcacc gtattgaaac tgagttaatg ggcaaatttg
     6001 atgaaggtaa actgcccacc gatccacacc tgatgctccg actggccatt gaaactgttg
     6061 ctcatgacta tgatgtcata gttattgaca gcgcgcctaa cctgggtatc ggcacgatta
     6121 atgtcgtatg tgctgctgat gtgctgattg ttcccacgcc tgctgagttg tttgactaca
     6181 cctccgcact gcagtttttc gatatgcttc gtgatctgct caagaacgtt gatcttaaag
     6241 ggttcgagcc tgatgtacgt attttgctta ccaaatacag caatagtaat ggctctcagt
     6301 ccccgtggat ggaggagcaa attcgggatg cctggggaag catggttcta aaaaatgttg
     6361 tacgtgaaac ggatgaagtt ggtaaaggtc agatccggat gagaactgtt tttgaacagg
     6421 ccattgatca acgcagttca actggtgcct ggagaaatgc tctttctatt tgggaacctg
     6481 tctgcaatga aattttcgat cgtctgatta aaccacgctg ggagattaga taatgaagcg
     6541 tgcgcctgtt attccaaaac atacgctcaa tactcaaccg gttgaagata cttcgttatc
     6601 gacaccagct gccccgatgg tggattcgtt aattgcgcgc gtaggagtaa tggctcgcgg
     6661 taatgccatt actttgcctg tatgtggtcg ggatgtgaag tttactcttg aagtgctccg
     6721 gggtgatagt gttgagaaaa cctctcgggt atggtcaggt aatgaacgtg accaggagct
     6781 gcttactgag gacgcactgg atgatctcat cccttctttt ctactgactg gtcaacagac
     6841 accggcgttc ggtcgaagag tatctggtgt catagaaatt gccgatggga gtcgccgtcg
     6901 taaagctgct gcacttaccg aaagtgatta tcgtgttctg gttggcgagc tggatgatga
     6961 gcagatggct gcattatcca gattgggtaa cgattatcgc ccaacaagtg cttatgaacg
     7021 tggtcagcgt tatgcaagcc gattgcagaa tgaatttgct ggaaatattt ctgcgctggc
     7081 tgatgcggaa aatatttcac gtaagattat tacccgctgt atcaacaccg ccaaattgcc
     7141 taaatcagtt gttgctcttt tttctcaccc cggtgaacta tctgcccggt caggtgatgc
     7201 acttcaaaaa gcctttacag ataaagagga attacttaag cagcaggcat ctaaccttca
     7261 tgagcagaaa aaagctgggg tgatatttga agctgaagaa gttatcactc ttttaacttc
     7321 tgtgcttaaa acgtcatctg catcaagaac tagtttaagc tcacgacatc agtttgctcc
     7381 tggagcgaca gtattgtata agggcgataa aatggtgctt aacctggaca ggtcacgtgt
     7441 tccaactgag tgtatagaga aaattgaggc cattcttaag gaacttgaaa agccagcacc
     7501 ctgatgcgac cacgttttag tctacgttta tctgtcttta cttaatgtcc tttgttacag
     7561 gccagaaagc ataactggcc tgaatattct ctctgggccc actgttccac ttgtatcgtc
     7621 ggtctgataa tcagactggg accacggtcc cactcgtatc gtcggtctga ttattagtct
     7681 gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact
     7741 cgtatcgtcg gtctgataat cagactggga ccacggtccc actcgtatcg tcggtctgat
     7801 tattagtctg ggaccatggt cccactcgta tcgtcggtct gattattagt ctgggaccac
     7861 ggtcccactc gtatcgtcgg tctgattatt agtctggaac cacggtccca ctcgtatcgt
     7921 cggtctgatt attagtctgg gaccacggtc ccactcgtat cgtcggtctg attattagtc
     7981 tgggaccacg atcccactcg tgttgtcggt ctgattatcg gtctg
//
