LOCUS       sequence-465763-        8660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOBK360_VR_N4 assembled plasmid (Kanamycin R) containing two 
            inserts (POC1544-POC1545) assembled in POC1520 using the 
            site-selective methylation protection approach.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8660)
  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 8660)
  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: POC1549
FEATURES             Location/Qualifiers
     source          1..8660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2353..2457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2458..3252
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      4067..4286
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             4377..5132
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    5135..5385
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5711..6886
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6886..7857
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    7930..8403
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcccag gaagaagttt cacctgtgat
       61 ttcctcttcc ccagagcccg tgctgggaaa taagacatgg gacagagaga ccagggactt
      121 gacagggaac ggaaaggacc tcaggatgac cctggctcat atcaaggacc agaaagaagg
      181 tgagagtcgg caggggcaag agtgactgga gaggcctttt ccagaaaagt taggggcaga
      241 gagcagggac ctgtctcttc ccactggatc tggctcaggc tgggggtgag gaatgggggt
      301 cagtggaact cagcagggag gtgagccggc actcagccca cacagggagg cgagtgggca
      361 gcagggaggg ctgcggcgcc tgctctgtcc ccatcccggt ctctgtgtct cttgaactca
      421 ctagggcgca tccaggtggg gtgagctggg aatcacgtgc tgaatgctaa gggcctggat
      481 gaaggcagct gtgaggccca gcctgggggc cctgcctgtg tagccctttg gagacccctt
      541 gtaacaggga gggtcctgag cacacatggc catctctgtc cactttgcac atgcacctcc
      601 tccaggagct ttcttggggt tgtcgtgtcc tctgcaccat tcgaggccct actctttctc
      661 cctgagccaa tttccctttc ctctccagga ggttctgtgt ggagatggtg gctgtggcag
      721 tggcagttcc caggtgcaga gggtgggcag aggcagcctc aggctaaggg gtctccccta
      781 ctccacatgg agaaaatcag gtcaagagat cgagaccatc ctgatcaaca tggtgaaacc
      841 ctgtctctac taaaaataca aaaaattagc tgggcctggt ggcgtgtgcc tgtagtccca
      901 gctactcagg agctgaggca catgcaccac catacccggc taatttttgc atttttcata
      961 gagacaggat tttgccatgt tggccaggct ggtctcaaac tcctgacctc aggtgatctg
     1021 cctgcctcag cctcccaaag tgctgggatt acaagcatga gccaccatac ccggcctatt
     1081 ttattacatt ttaaactgat gcacttcttg gtcaaggcaa cctcagcccc acccacgtaa
     1141 ggtggtcatg gcagagagtg taagggtgac acctgtgaaa aagacccaag gcagggatgg
     1201 gagcccttct tgcagcagga gtggatgcag gacctgcctg gaagcaagag aaggacgagg
     1261 gaccctggct gggcccgtat ttttagtaga gacagtttct ccgtattggt caggctggtc
     1321 tcgaactcct gacctcaggt gattgcctca cagttcaagt caggcttggg gaaagtagtg
     1381 agaccctgtc tccagccaaa aaaaaaaaat taaattaaaa tatactgtac tataatagtg
     1441 ttaattgtaa gcacagtgta gtgcaaccga tctttagaat atttcaccta ctggccggtg
     1501 ctgggtttca gcatcctgca atttcagaac tactgtgaat acaaaagaaa gaaaggtcct
     1561 gcccaggatg ggagtcactc taattgccat tgagaaaaat atgcctaggc cagaagattg
     1621 gtgatggtgg gaatagcagg gacatagttt agacaaggtc attgagcaag acgtttctga
     1681 gaacttgacc actggttatc tgagaagaga cctgaatcgt gtgaggagtg agtcatgtga
     1741 atccctgggg agcaggtaca tgtgggagtc agagcatggg taggtgggat gttcaaagtg
     1801 aagttgacat ctacaataag tagtaagtgt agacctcaga aatttaacaa gattctactt
     1861 tttcccccaa aaaaatgtcc tcacaatttc ctgctgaaag gagagggttc attcagggga
     1921 ccacatgatg ccgttcttct gctgcagttc taaattaagt tgtccttccc tctaactaca
     1981 ggtctcacaa acccttgcag ccagcttcaa tgattttagc ttgactgttt ttgacttggg
     2041 tattttgcca aaatggtatg tttttctata gaatttgacg ttgtggagac cctgcttttc
     2101 tgccaggtcc ccagagggga tacagtgctt tgtacggcaa tgacattccc ctgagctata
     2161 tccctggtta ctacagcatt tcctgatctt gcctaacagg attacttccc tgaactgtaa
     2221 aattcctgca gcattgtata cagagaaaga ataggagaca tggcaatcat ggacaagaaa
     2281 ggaggaaatc gtgtgagact gagaccaagt cgtgagaggt actgcaccac gaagtgtgcg
     2341 gccgccttcg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2401 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2461 gctaaaatga gaatatcacc ggaattgaaa aaactgatcg aaaaataccg ctgcgtaaaa
     2521 gatacggaag gaatgtctcc tgctaaggta tataagctgg tgggagaaaa tgaaaaccta
     2581 tatttaaaaa tgacggacag ccggtataaa gggaccacct atgatgtgga acgggaaaag
     2641 gacatgatgc tatggctgga aggaaagctg cctgttccaa aggtcctgca ctttgaacgg
     2701 catgatggct ggagcaatct gctcatgagt gaggccgatg gcgtcctttg ctcggaagag
     2761 tatgaagatg aacaaagccc tgaaaagatt atcgagctgt atgcggagtg catcaggctc
     2821 tttcactcca tcgacatatc ggattgtccc tatacgaata gcttagacag ccgcttagcc
     2881 gaattggatt acttactgaa taacgatctg gccgatgtgg attgcgaaaa ctgggaagag
     2941 gacactccat ttaaagatcc gcgcgagctg tatgattttt taaagacgga aaagcccgaa
     3001 gaggaacttg tcttttccca cggcgacctg ggagacagca acatctttgt gaaagatggc
     3061 aaagtaagtg gctttattga tcttgggaga agcggcaggg cggacaagtg gtatgacatt
     3121 gccttctgcg tccggtcgct cagggaggat atcggggaag aacagtatgt cgagctattt
     3181 tttgacttac tggggatcaa gcctgattgg gagaaaataa aatattatat tttactggat
     3241 gaattgtttt aggagctgga gcggcgtaac cgtcgcacag gaaggacaga gaaagcgcgg
     3301 atctgggaag tgacggacag aacggtcagg acctggattg gggaggcggt tgccgccgct
     3361 gctgctgacg gtgtgacgtt ctctgttccg gtcacaccac atacgttccg ccattcctat
     3421 gcgatgcaca tgctgtatgc cggtataccg ctgaaagttc tgcaaagcct gatgggacat
     3481 aagtccatca gttcaacgga agtctacacg aaggtttttg cgctggatgt ggctgcccgg
     3541 ctccgggtgc agtttgcgat gccggagtct gatgcggttg cgatgctgaa acaattatcc
     3601 tgagaataaa tgccttggcc tttatatgga aatgtggaac tgagtggata tgctgttttt
     3661 gtctgttaaa cagagaagct ggctgttatc cactgagaag cgaacgaaac agtcgggaaa
     3721 atctcccatt atcgtagaga tccgcattat taatctcagg agcctgtgta gcgtttatag
     3781 gaagtagtgt tctgtcatga tgcctgcaag cggtaacgaa aacgatttga atatgccttc
     3841 aggaacaata gaaatcttcg tgcggtgtta cgttgaagtg gagcggatta tgtcagcaat
     3901 ggacagaaca acctaatgaa cacagaacca tgatgtggtc tgtcctttta cagccagtag
     3961 tgctcgccgc agtcgagcga cagggcgaag ccctcgagtg agcgaggaag caccagggaa
     4021 cagcacttat atattctgct tacacacgat gcctgaaaaa acttcccttg gggttatcca
     4081 cttatccacg gggatatttt tataattatt ttttttatag tttttagatc ttctttttta
     4141 gagcgccttg taggccttta tccatgctgg ttctagagaa ggtgttgtga caaattgccc
     4201 tttcagtgtg acaaatcacc ctcaaatgac agtcctgtct gtgacaaatt gcccttaacc
     4261 ctgtgacaaa ttgccctcag aagaagctgt tttttcacaa agttatccct gcttattgac
     4321 tcttttttat ttagtgtgac aatctaaaaa cttgtcacac ttcacatgga tctgtcatgg
     4381 cggaaacagc ggttatcaat cacaagaaac gtaaaaatag cccgcgaatc gtccagtcaa
     4441 acgacctcac tgaggcggca tatagtctct cccgggatca aaaacgtatg ctgtatctgt
     4501 tcgttgacca gatcagaaaa tctgatggca ccctacagga acatgacggt atctgcgaga
     4561 tccatgttgc taaatatgct gaaatattcg gattgacctc tgcggaagcc agtaaggata
     4621 tacggcaggc attgaagagt ttcgcgggga aggaagtggt tttttatcgc cctgaagagg
     4681 atgccggcga tgaaaaaggc tatgaatctt ttccttggtt tatcaaacgt gcgcacagtc
     4741 catccagagg gctttacagt gtacatatca acccatatct cattcccttc tttatcgggt
     4801 tacagaaccg gtttacgcag tttcggctta gtgaaacaaa agaaatcacc aatccgtatg
     4861 ccatgcgttt atacgaatcc ctgtgtcagt atcgtaagcc ggatggctca ggcatcgtgt
     4921 ctctgaaaat cgactggatc atagagcgtt accagctgcc tcaaagttac cagcgtatgc
     4981 ctgacttccg ccgccgcttc ctgcaggtct gtgttaatga gatcaacagc agaactccaa
     5041 tgcgcctctc atacattgag aaaaagaaag gccgccagac gactcatatc gtattttcct
     5101 tccgcgatat cacttccatg acgacaggat agtctgaggg ttatctgtca cagatttgag
     5161 ggtggttcgt cacatttgtt ctgacctact gagggtaatt tgtcacagtt ttgctgtttc
     5221 cttcagcctg catggatttt ctcatacttt ttgaactgta atttttaagg aagccaaatt
     5281 tgagggcagt ttgtcacagt tgatttcctt ctctttccct tcgtcatgtg acctgatatc
     5341 gggggttagt tcgtcatcat tgatgagggt tgattatcac agtttattac tctgaattgg
     5401 ctatccgcgt gtgtacctct acctggagtt tttcccacgg tggatatttc ttcttgcgct
     5461 gagcgtaaga gctatctgac agaacagttc ttctttgctt cctcgccagt tcgctcgcta
     5521 tgctcggtta cacggctgcg gcgagcgcta gtgataataa gtgactgagg tatgtggtgt
     5581 tcttatctcc ttttgtagtg ttgctcttat tttaaacaac tttgcggttt tttgatgact
     5641 ttgcgatttt gttgttgctt tgcagtaaat tgcaagattt aataaaaaaa cgcaaagcaa
     5701 tgattaaagg atgttcagaa tgaaactcat ggaaacactt aaccagtgca taaacgctgg
     5761 tcatgaaatg acgaaggcta tcgccattgc acagtttaat gatgacagcc cggaagcgag
     5821 gaaaataacc cggcgctgga gaataggtga agcagcggat ttagttgggg tttcttctca
     5881 ggctatcaga gatgccgaga aagcagggcg actaccgcac ccggatatgg aaattcgagg
     5941 acgggttgag caacgtgttg gttatacaat tgaacaaatt aatcatatgc gtgatgtgtt
     6001 tggtacgcga ttgcgacgtg ctgaggacgt atttccaccg gtgatcgggg ttgctgccca
     6061 taaaggtggc gtttacaaaa cctcagtttc tgttcatctt gctcaggatc tggctctgaa
     6121 ggggctacgt gttttgctcg tggaaggtaa cgacccccag ggaacagcct caatgtatca
     6181 cggatgggta ccagatcttc atattcatgc agaagatact ctcctgcctt tctatcttgg
     6241 ggaaaaggac gatgtcactt atgcaataaa gcccacttgc tggccggggc ttgacattat
     6301 tccttcctgt ctggctctgc accgtattga aactgagtta atgggcaaat ttgatgaagg
     6361 taaactgccc accgatccac acctgatgct ccgactggcc attgaaactg ttgctcatga
     6421 ctatgatgtc atagttattg acagcgcgcc taacctgggt atcggcacga ttaatgtcgt
     6481 atgtgctgct gatgtgctga ttgttcccac gcctgctgag ttgtttgact acacctccgc
     6541 actgcagttt ttcgatatgc ttcgtgatct gctcaagaac gttgatctta aagggttcga
     6601 gcctgatgta cgtattttgc ttaccaaata cagcaatagt aatggctctc agtccccgtg
     6661 gatggaggag caaattcggg atgcctgggg aagcatggtt ctaaaaaatg ttgtacgtga
     6721 aacggatgaa gttggtaaag gtcagatccg gatgagaact gtttttgaac aggccattga
     6781 tcaacgcagt tcaactggtg cctggagaaa tgctctttct atttgggaac ctgtctgcaa
     6841 tgaaattttc gatcgtctga ttaaaccacg ctgggagatt agataatgaa gcgtgcgcct
     6901 gttattccaa aacatacgct caatactcaa ccggttgaag atacttcgtt atcgacacca
     6961 gctgccccga tggtggattc gttaattgcg cgcgtaggag taatggctcg cggtaatgcc
     7021 attactttgc ctgtatgtgg tcgggatgtg aagtttactc ttgaagtgct ccggggtgat
     7081 agtgttgaga aaacctctcg ggtatggtca ggtaatgaac gtgaccagga gctgcttact
     7141 gaggacgcac tggatgatct catcccttct tttctactga ctggtcaaca gacaccggcg
     7201 ttcggtcgaa gagtatctgg tgtcatagaa attgccgatg ggagtcgccg tcgtaaagct
     7261 gctgcactta ccgaaagtga ttatcgtgtt ctggttggcg agctggatga tgagcagatg
     7321 gctgcattat ccagattggg taacgattat cgcccaacaa gtgcttatga acgtggtcag
     7381 cgttatgcaa gccgattgca gaatgaattt gctggaaata tttctgcgct ggctgatgcg
     7441 gaaaatattt cacgtaagat tattacccgc tgtatcaaca ccgccaaatt gcctaaatca
     7501 gttgttgctc ttttttctca ccccggtgaa ctatctgccc ggtcaggtga tgcacttcaa
     7561 aaagccttta cagataaaga ggaattactt aagcagcagg catctaacct tcatgagcag
     7621 aaaaaagctg gggtgatatt tgaagctgaa gaagttatca ctcttttaac ttctgtgctt
     7681 aaaacgtcat ctgcatcaag aactagttta agctcacgac atcagtttgc tcctggagcg
     7741 acagtattgt ataagggcga taaaatggtg cttaacctgg acaggtcacg tgttccaact
     7801 gagtgtatag agaaaattga ggccattctt aaggaacttg aaaagccagc accctgatgc
     7861 gaccacgttt tagtctacgt ttatctgtct ttacttaatg tcctttgtta caggccagaa
     7921 agcataactg gcctgaatat tctctctggg cccactgttc cacttgtatc gtcggtctga
     7981 taatcagact gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca
     8041 cggtcccact cgtatcgtcg gtctgattat tagtctggga ccacggtccc actcgtatcg
     8101 tcggtctgat aatcagactg ggaccacggt cccactcgta tcgtcggtct gattattagt
     8161 ctgggaccat ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacggtccca
     8221 ctcgtatcgt cggtctgatt attagtctgg aaccacggtc ccactcgtat cgtcggtctg
     8281 attattagtc tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc
     8341 acgatcccac tcgtgttgtc ggtctgatta tcggtctggg accacggtcc cacttgtatt
     8401 gtcgatcaga ctatcagcgt gagactacga ttccatcaat gcctgtcaag ggcaagtatt
     8461 gacatgtcgt cgtaacctgt agaacggagt aacctcggtg tgcggttgta tgcctgctgt
     8521 ggattgctgc tgtgtcctgc ttatccacaa cattttgcgc acggttatgt ggacaaaata
     8581 cctggttacc caggccgtgc cggcacgtta accgggctgc atccgatgca agtgtgtcgc
     8641 tgtcgacgca agcggccgca
//
