LOCUS       sequence-469009-        9890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOBK360_VL_N1 assembled plasmid (Kanamycin R) containing four 
            inserts (POC1535-POC1538) assembled in POC1518 using the 
            site-selective methylation protection approach.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9890)
  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 9890)
  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: POC1546
FEATURES             Location/Qualifiers
     source          1..9890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3583..3687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3688..4482
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      5297..5516
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             5607..6362
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    6365..6615
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             6941..8116
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             8116..9087
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    9160..9633
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcactc catctataaa caggtgtttc
       61 ccagtctgaa agggtgtttc atttaaaaca ggtctaactt ctgtttgata actaattaaa
      121 tctaaacatc tgtgctctgg ggatggcttc atactttaaa attcttgaat cagtaacatc
      181 tggccagcta tttgtgacga agtgaagctt taacattcct tgtccaagtg ggtctcctcc
      241 atctaaacca gcatatttcc tcatttgttc cttaagcctg ttaaaaaatt ccacatattc
      301 tgggtgcagg gtaacgattc tctaattcct tttattatta tttcttgcag gtctctcata
      361 ttcctcctat gagctatgtt gttattatcc cactgggaat cttgagtagg aaatttctgt
      421 ttggctgcag ggacgttttg accaggagga tgataacctc ttgccattct tttaccttct
      481 gtctctatgg attgtggtgc tgtttacatt ccagattttt attaatatgt gaggctttgt
      541 gccgagacta gctcagttgg gaagacccta acccagtggc actagaggaa tcaaagacac
      601 acacacagaa atatagaggt gtgaagtggg aaatcaggga tctcacagcc ttcagagctg
      661 acagccttga acagagattt acccccatat ttattaactc aagccagtga taagcattgt
      721 ttctatagat tatagattaa ctaaaagtat tccttatggg aaacaaacgg atgggccaaa
      781 ataaagggat gggtttggct agttatctgc agcaggagca tgtccttaag gcacagatca
      841 ctcatgctat tgtttgtggt ttaagaacga ctttaagcag ttttccgccc tgggtgggcc
      901 aggtgtcatt atggtttggt ggatttctgt agtaggacca tttgagacct ttctcttcct
      961 cttttgtgtg attgctttac cagtgagttt tatacttgtg tatgttttca tggtgataat
     1021 gtggaaatgt tgaaaatgtc gtttcaattc caggtttagg actttcttga gcatttcctg
     1081 tagactcttg gcttggcagt tcttttcctt caggtttctg ctaagaaatc tgctgttaga
     1141 gctgggcaca gtggctcaca cctgtaatcc cagcactttg ggaggctgag gtgggtggat
     1201 catgatgtca agagattgag accatcctgg ccaacttggt gaaatcccgt ctctactaaa
     1261 aatacaaaaa aaaaaaaaaa aattagctgt gtctagtatg ttgttgaagc tttcaaatgt
     1321 atttggaatt tcatgtcatg aattctttaa ttccagaatt tctgtttttt tttttaaatc
     1381 tatatctttt gtaaacttct tatttgtatc ctttgcggct tttttttgga gcttacaggt
     1441 cttaggtaga tttaaagatt ctctgatttg tgatagataa ggaagcaaag gagaggaatg
     1501 ttcaggcctg gcctgtgggc ttgactctct ccaggtccct caggaagaaa tttagaacaa
     1561 agaacggcgg tcagagttca gttctcagtt tcccctttat aaggtctccc tgtcagtgga
     1621 tctattagga gggaatccat gtttctgaaa aacaactcag ggacatatgt taagatgtta
     1681 tttttagttt ctatagagaa tgaaacatct tgtgactcta acttccttgg ctattgtttc
     1741 aagctatcat tccttcttgc ttataaggtc actcacttac tttttagggc tggctaggtg
     1801 cctggaattt ctcttgaagg gactgaaggt ttttctttat ttctagattg ggaggcccca
     1861 gcaggcttct aaaagaggtc cctgctttat ctcagatgca aatgctcgag tgttacaaaa
     1921 gaacttgaat gggaggtact gcaaccatgt ggaccaccga gtcatatttc tttacaccag
     1981 aaaatgcacc tgcatatcct cctgtttatg gattgcactg ggataattgt taatgaccaa
     2041 agaggattct ggtaatgtgc caggatcagg agatctgtga ttggccaggc acagtggctc
     2101 acacttataa ttccatcagt ttgggaggca atggtaggaa cattgtttaa gtccaggagt
     2161 ttgagaccag actgggcaga atagtgagac ccattcctac aaaatattta aaaattagtt
     2221 gggcatgttg gtgtgcactt gtctccgtgg ggacagaggc tcttgtgctc agagcccttc
     2281 caggcctcac tgtgcacctc ttcatctggc tgctcatttg tatcctttat aactgctatg
     2341 gtttgaatgt ttcccccaaa tgctacaatg atttgtgtct ctgtttccaa ggtgcatctt
     2401 cagcttggtg aaataaactt ctaaactgac tgagacctgt ctcagacgtt ttttggttta
     2461 catggctata gcaacttcct gacagagacc aacaatgagt gaccacagcc catgacacag
     2521 ccctcaggag accctgagaa catgtgctca aggtacattt aaggaatatg ggggttcctt
     2581 gtctgagtgt cctctagacg tgggaccaga actgtgcagt ctgctctggg tgtggtctga
     2641 tcacacccct tagaactggt ccagggttca gtccttgtgc tcattctttt ccatagtcag
     2701 tcactccctt ggtgcttcat ccatgcctga ggttttaagt ctcatatata tggtgtgacc
     2761 tcctaaatct atttctccag ccacagcagc tccactagaa ccagggcagg gagagagggc
     2821 agaagagcag agcagaaaag gagctctaga agcagagcag gaggtgaatg gctctgaaaa
     2881 tttgtctcag aatgcacaga gacccccgtg tgcaggggcc gccctgggcg atgtgtgagc
     2941 ctctgtggtc acagctccca ctggacaagt ttccactgaa gggacaagga caatggagca
     3001 gtgaaggtga cccagctgag gactaaccac ataaagccca tgatggactc aacaccaaat
     3061 gggcacaggc cccatccatt tggattggaa agttcgagtc tctgatccag gagcgcaggt
     3121 cctcgttcag ggcgatgtaa tccttgccgt cgtaggcgta ctggtcatgc ccaatttact
     3181 agatatatat taatactacg tcgcagccgt acatgctctg gagggtgtga gaccctggcc
     3241 ccggccccgc ggaattaata tttgatatat cttaaaatat gtatataaat tttactagta
     3301 tatatacttt aaatttaaat ataaattaaa tttatataac acggaggtgt agaaatacct
     3361 catggagtgg gagcctgggg gtgaggaggg gctgagaccc gcccgaccct cctaatttaa
     3421 atactcctca cgcataggga ttctcataga gaccagtttc ctttttgttt gttaatacag
     3481 taggtagcac actaatgtca tttcttcact tgcaatggtt gggttttggt gagaccaagt
     3541 cgtgagaggt actgcaccac gaagtgtgcg gccgccttcg tgcgcggaac ccctatttgt
     3601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3661 cttcaataat attgaaaaag gaagagtatg gctaaaatga gaatatcacc ggaattgaaa
     3721 aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta
     3781 tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa
     3841 gggaccacct atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg
     3901 cctgttccaa aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt
     3961 gaggccgatg gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt
     4021 atcgagctgt atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc
     4081 tatacgaata gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg
     4141 gccgatgtgg attgcgaaaa ctgggaagag gacactccat ttaaagatcc gcgcgagctg
     4201 tatgattttt taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg
     4261 ggagacagca acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga
     4321 agcggcaggg cggacaagtg gtatgacatt gccttctgcg tccggtcgct cagggaggat
     4381 atcggggaag aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg
     4441 gagaaaataa aatattatat tttactggat gaattgtttt aggagctgga gcggcgtaac
     4501 cgtcgcacag gaaggacaga gaaagcgcgg atctgggaag tgacggacag aacggtcagg
     4561 acctggattg gggaggcggt tgccgccgct gctgctgacg gtgtgacgtt ctctgttccg
     4621 gtcacaccac atacgttccg ccattcctat gcgatgcaca tgctgtatgc cggtataccg
     4681 ctgaaagttc tgcaaagcct gatgggacat aagtccatca gttcaacgga agtctacacg
     4741 aaggtttttg cgctggatgt ggctgcccgg ctccgggtgc agtttgcgat gccggagtct
     4801 gatgcggttg cgatgctgaa acaattatcc tgagaataaa tgccttggcc tttatatgga
     4861 aatgtggaac tgagtggata tgctgttttt gtctgttaaa cagagaagct ggctgttatc
     4921 cactgagaag cgaacgaaac agtcgggaaa atctcccatt atcgtagaga tccgcattat
     4981 taatctcagg agcctgtgta gcgtttatag gaagtagtgt tctgtcatga tgcctgcaag
     5041 cggtaacgaa aacgatttga atatgccttc aggaacaata gaaatcttcg tgcggtgtta
     5101 cgttgaagtg gagcggatta tgtcagcaat ggacagaaca acctaatgaa cacagaacca
     5161 tgatgtggtc tgtcctttta cagccagtag tgctcgccgc agtcgagcga cagggcgaag
     5221 ccctcgagtg agcgaggaag caccagggaa cagcacttat atattctgct tacacacgat
     5281 gcctgaaaaa acttcccttg gggttatcca cttatccacg gggatatttt tataattatt
     5341 ttttttatag tttttagatc ttctttttta gagcgccttg taggccttta tccatgctgg
     5401 ttctagagaa ggtgttgtga caaattgccc tttcagtgtg acaaatcacc ctcaaatgac
     5461 agtcctgtct gtgacaaatt gcccttaacc ctgtgacaaa ttgccctcag aagaagctgt
     5521 tttttcacaa agttatccct gcttattgac tcttttttat ttagtgtgac aatctaaaaa
     5581 cttgtcacac ttcacatgga tctgtcatgg cggaaacagc ggttatcaat cacaagaaac
     5641 gtaaaaatag cccgcgaatc gtccagtcaa acgacctcac tgaggcggca tatagtctct
     5701 cccgggatca aaaacgtatg ctgtatctgt tcgttgacca gatcagaaaa tctgatggca
     5761 ccctacagga acatgacggt atctgcgaga tccatgttgc taaatatgct gaaatattcg
     5821 gattgacctc tgcggaagcc agtaaggata tacggcaggc attgaagagt ttcgcgggga
     5881 aggaagtggt tttttatcgc cctgaagagg atgccggcga tgaaaaaggc tatgaatctt
     5941 ttccttggtt tatcaaacgt gcgcacagtc catccagagg gctttacagt gtacatatca
     6001 acccatatct cattcccttc tttatcgggt tacagaaccg gtttacgcag tttcggctta
     6061 gtgaaacaaa agaaatcacc aatccgtatg ccatgcgttt atacgaatcc ctgtgtcagt
     6121 atcgtaagcc ggatggctca ggcatcgtgt ctctgaaaat cgactggatc atagagcgtt
     6181 accagctgcc tcaaagttac cagcgtatgc ctgacttccg ccgccgcttc ctgcaggtct
     6241 gtgttaatga gatcaacagc agaactccaa tgcgcctctc atacattgag aaaaagaaag
     6301 gccgccagac gactcatatc gtattttcct tccgcgatat cacttccatg acgacaggat
     6361 agtctgaggg ttatctgtca cagatttgag ggtggttcgt cacatttgtt ctgacctact
     6421 gagggtaatt tgtcacagtt ttgctgtttc cttcagcctg catggatttt ctcatacttt
     6481 ttgaactgta atttttaagg aagccaaatt tgagggcagt ttgtcacagt tgatttcctt
     6541 ctctttccct tcgtcatgtg acctgatatc gggggttagt tcgtcatcat tgatgagggt
     6601 tgattatcac agtttattac tctgaattgg ctatccgcgt gtgtacctct acctggagtt
     6661 tttcccacgg tggatatttc ttcttgcgct gagcgtaaga gctatctgac agaacagttc
     6721 ttctttgctt cctcgccagt tcgctcgcta tgctcggtta cacggctgcg gcgagcgcta
     6781 gtgataataa gtgactgagg tatgtggtgt tcttatctcc ttttgtagtg ttgctcttat
     6841 tttaaacaac tttgcggttt tttgatgact ttgcgatttt gttgttgctt tgcagtaaat
     6901 tgcaagattt aataaaaaaa cgcaaagcaa tgattaaagg atgttcagaa tgaaactcat
     6961 ggaaacactt aaccagtgca taaacgctgg tcatgaaatg acgaaggcta tcgccattgc
     7021 acagtttaat gatgacagcc cggaagcgag gaaaataacc cggcgctgga gaataggtga
     7081 agcagcggat ttagttgggg tttcttctca ggctatcaga gatgccgaga aagcagggcg
     7141 actaccgcac ccggatatgg aaattcgagg acgggttgag caacgtgttg gttatacaat
     7201 tgaacaaatt aatcatatgc gtgatgtgtt tggtacgcga ttgcgacgtg ctgaggacgt
     7261 atttccaccg gtgatcgggg ttgctgccca taaaggtggc gtttacaaaa cctcagtttc
     7321 tgttcatctt gctcaggatc tggctctgaa ggggctacgt gttttgctcg tggaaggtaa
     7381 cgacccccag ggaacagcct caatgtatca cggatgggta ccagatcttc atattcatgc
     7441 agaagatact ctcctgcctt tctatcttgg ggaaaaggac gatgtcactt atgcaataaa
     7501 gcccacttgc tggccggggc ttgacattat tccttcctgt ctggctctgc accgtattga
     7561 aactgagtta atgggcaaat ttgatgaagg taaactgccc accgatccac acctgatgct
     7621 ccgactggcc attgaaactg ttgctcatga ctatgatgtc atagttattg acagcgcgcc
     7681 taacctgggt atcggcacga ttaatgtcgt atgtgctgct gatgtgctga ttgttcccac
     7741 gcctgctgag ttgtttgact acacctccgc actgcagttt ttcgatatgc ttcgtgatct
     7801 gctcaagaac gttgatctta aagggttcga gcctgatgta cgtattttgc ttaccaaata
     7861 cagcaatagt aatggctctc agtccccgtg gatggaggag caaattcggg atgcctgggg
     7921 aagcatggtt ctaaaaaatg ttgtacgtga aacggatgaa gttggtaaag gtcagatccg
     7981 gatgagaact gtttttgaac aggccattga tcaacgcagt tcaactggtg cctggagaaa
     8041 tgctctttct atttgggaac ctgtctgcaa tgaaattttc gatcgtctga ttaaaccacg
     8101 ctgggagatt agataatgaa gcgtgcgcct gttattccaa aacatacgct caatactcaa
     8161 ccggttgaag atacttcgtt atcgacacca gctgccccga tggtggattc gttaattgcg
     8221 cgcgtaggag taatggctcg cggtaatgcc attactttgc ctgtatgtgg tcgggatgtg
     8281 aagtttactc ttgaagtgct ccggggtgat agtgttgaga aaacctctcg ggtatggtca
     8341 ggtaatgaac gtgaccagga gctgcttact gaggacgcac tggatgatct catcccttct
     8401 tttctactga ctggtcaaca gacaccggcg ttcggtcgaa gagtatctgg tgtcatagaa
     8461 attgccgatg ggagtcgccg tcgtaaagct gctgcactta ccgaaagtga ttatcgtgtt
     8521 ctggttggcg agctggatga tgagcagatg gctgcattat ccagattggg taacgattat
     8581 cgcccaacaa gtgcttatga acgtggtcag cgttatgcaa gccgattgca gaatgaattt
     8641 gctggaaata tttctgcgct ggctgatgcg gaaaatattt cacgtaagat tattacccgc
     8701 tgtatcaaca ccgccaaatt gcctaaatca gttgttgctc ttttttctca ccccggtgaa
     8761 ctatctgccc ggtcaggtga tgcacttcaa aaagccttta cagataaaga ggaattactt
     8821 aagcagcagg catctaacct tcatgagcag aaaaaagctg gggtgatatt tgaagctgaa
     8881 gaagttatca ctcttttaac ttctgtgctt aaaacgtcat ctgcatcaag aactagttta
     8941 agctcacgac atcagtttgc tcctggagcg acagtattgt ataagggcga taaaatggtg
     9001 cttaacctgg acaggtcacg tgttccaact gagtgtatag agaaaattga ggccattctt
     9061 aaggaacttg aaaagccagc accctgatgc gaccacgttt tagtctacgt ttatctgtct
     9121 ttacttaatg tcctttgtta caggccagaa agcataactg gcctgaatat tctctctggg
     9181 cccactgttc cacttgtatc gtcggtctga taatcagact gggaccacgg tcccactcgt
     9241 atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg gtctgattat
     9301 tagtctggga ccacggtccc actcgtatcg tcggtctgat aatcagactg ggaccacggt
     9361 cccactcgta tcgtcggtct gattattagt ctgggaccat ggtcccactc gtatcgtcgg
     9421 tctgattatt agtctgggac cacggtccca ctcgtatcgt cggtctgatt attagtctgg
     9481 aaccacggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg
     9541 tatcgtcggt ctgattatta gtctgggacc acgatcccac tcgtgttgtc ggtctgatta
     9601 tcggtctggg accacggtcc cacttgtatt gtcgatcaga ctatcagcgt gagactacga
     9661 ttccatcaat gcctgtcaag ggcaagtatt gacatgtcgt cgtaacctgt agaacggagt
     9721 aacctcggtg tgcggttgta tgcctgctgt ggattgctgc tgtgtcctgc ttatccacaa
     9781 cattttgcgc acggttatgt ggacaaaata cctggttacc caggccgtgc cggcacgtta
     9841 accgggctgc atccgatgca agtgtgtcgc tgtcgacgca agcggccgca
//
