LOCUS       sequence-225176-        8069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12405, MetClo assembly vector with F replication origin and 
            ampicillin resistance for BsaI-based MetClo, adaptor sequence type 
            b-c.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  AUTHORS   Lin D, O'Callaghan CA
  TITLE     MetClo: methylase-assisted hierarchical DNA assembly using a single 
            type IIS restriction enzyme.
  JOURNAL   Nucleic Acids Res. 2018 Jul 9. pii: 5050631. doi: 
            10.1093/nar/gky596.
  PUBMED    29986052
REFERENCE   2  (bases 1 to 8069)
  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. 2018 Jul 9. pii: 5050631. doi: 10.1093/nar/gky596."
COMMENT     Sequence Label: pMXBP_bFc
FEATURES             Location/Qualifiers
     source          1..8069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      224..812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     713..732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     966..983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1100..1121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1136..1141,1142..1159,1160..1166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1174..1190
                     /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)."
     primer_bind     1179..1201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1198..1214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1198..1214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1210..1533
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    1227..1283
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1284..1301)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1284..1300)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1293..1315)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1509..1528)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1696..1800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1801..1869,1870..2661)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2019..2038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3476..3695
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3786..4541
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4544..4794
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5120..6295
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6295..7266
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    7339..7812
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
ORIGIN
        1 cactctgtga cttggtctca tgccggagtg agaccggcct agcaggtgtg agtttgtcag
       61 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
      121 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
      181 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
      241 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      301 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      361 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      421 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      481 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      541 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      601 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      661 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      721 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      781 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      841 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      901 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      961 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     1021 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     1081 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     1141 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     1201 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat
     1261 ccccgggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1321 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1381 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1441 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     1501 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     1561 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1621 acaagtacca cctggaacgc caaaggtctc acgctacagt gagaccaagt cacgaagtgt
     1681 gcggccgcct tcgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1741 tatccgctca tgagacaata accttgataa atgcttcaat aatattgaaa aaggaagagt
     1801 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1861 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1921 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1981 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2041 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2101 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2161 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2221 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2281 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2341 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2401 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2461 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggttct
     2521 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2581 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2641 tcactgatta agcattggta agagctggag cggcgtaacc gtcgcacagg aaggacagag
     2701 aaagcgcgga tctgggaagt gacggacaga acggtcagga cctggattgg ggaggcggtt
     2761 gccgccgctg ctgctgacgg tgtgacgttc tctgttccgg tcacaccaca tacgttccgc
     2821 cattcctatg cgatgcacat gctgtatgcc ggtataccgc tgaaagttct gcaaagcctg
     2881 atgggacata agtccatcag ttcaacggaa gtctacacga aggtttttgc gctggatgtg
     2941 gctgcccggc tccgggtgca gtttgcgatg ccggagtctg atgcggttgc gatgctgaaa
     3001 caattatcct gagaataaat gccttggcct ttatatggaa atgtggaact gagtggatat
     3061 gctgtttttg tctgttaaac agagaagctg gctgttatcc actgagaagc gaacgaaaca
     3121 gtcgggaaaa tctcccatta tcgtagagat ccgcattatt aatctcagga gcctgtgtag
     3181 cgtttatagg aagtagtgtt ctgtcatgat gcctgcaagc ggtaacgaaa acgatttgaa
     3241 tatgccttca ggaacaatag aaatcttcgt gcggtgttac gttgaagtgg agcggattat
     3301 gtcagcaatg gacagaacaa cctaatgaac acagaaccat gatgtggtct gtccttttac
     3361 agccagtagt gctcgccgca gtcgagcgac agggcgaagc cctcgagtga gcgaggaagc
     3421 accagggaac agcacttata tattctgctt acacacgatg cctgaaaaaa cttcccttgg
     3481 ggttatccac ttatccacgg ggatattttt ataattattt tttttatagt ttttagatct
     3541 tcttttttag agcgccttgt aggcctttat ccatgctggt tctagagaag gtgttgtgac
     3601 aaattgccct ttcagtgtga caaatcaccc tcaaatgaca gtcctgtctg tgacaaattg
     3661 cccttaaccc tgtgacaaat tgccctcaga agaagctgtt ttttcacaaa gttatccctg
     3721 cttattgact cttttttatt tagtgtgaca atctaaaaac ttgtcacact tcacatggat
     3781 ctgtcatggc ggaaacagcg gttatcaatc acaagaaacg taaaaatagc ccgcgaatcg
     3841 tccagtcaaa cgacctcact gaggcggcat atagtctctc ccgggatcaa aaacgtatgc
     3901 tgtatctgtt cgttgaccag atcagaaaat ctgatggcac cctacaggaa catgacggta
     3961 tctgcgagat ccatgttgct aaatatgctg aaatattcgg attgacctct gcggaagcca
     4021 gtaaggatat acggcaggca ttgaagagtt tcgcggggaa ggaagtggtt ttttatcgcc
     4081 ctgaagagga tgccggcgat gaaaaaggct atgaatcttt tccttggttt atcaaacgtg
     4141 cgcacagtcc atccagaggg ctttacagtg tacatatcaa cccatatctc attcccttct
     4201 ttatcgggtt acagaaccgg tttacgcagt ttcggcttag tgaaacaaaa gaaatcacca
     4261 atccgtatgc catgcgttta tacgaatccc tgtgtcagta tcgtaagccg gatggctcag
     4321 gcatcgtgtc tctgaaaatc gactggatca tagagcgtta ccagctgcct caaagttacc
     4381 agcgtatgcc tgacttccgc cgccgcttcc tgcaggtctg tgttaatgag atcaacagca
     4441 gaactccaat gcgcctctca tacattgaga aaaagaaagg ccgccagacg actcatatcg
     4501 tattttcctt ccgcgatatc acttccatga cgacaggata gtctgagggt tatctgtcac
     4561 agatttgagg gtggttcgtc acatttgttc tgacctactg agggtaattt gtcacagttt
     4621 tgctgtttcc ttcagcctgc atggattttc tcatactttt tgaactgtaa tttttaagga
     4681 agccaaattt gagggcagtt tgtcacagtt gatttccttc tctttccctt cgtcatgtga
     4741 cctgatatcg ggggttagtt cgtcatcatt gatgagggtt gattatcaca gtttattact
     4801 ctgaattggc tatccgcgtg tgtacctcta cctggagttt ttcccacggt ggatatttct
     4861 tcttgcgctg agcgtaagag ctatctgaca gaacagttct tctttgcttc ctcgccagtt
     4921 cgctcgctat gctcggttac acggctgcgg cgagcgctag tgataataag tgactgaggt
     4981 atgtggtgtt cttatctcct tttgtagtgt tgctcttatt ttaaacaact ttgcggtttt
     5041 ttgatgactt tgcgattttg ttgttgcttt gcagtaaatt gcaagattta ataaaaaaac
     5101 gcaaagcaat gattaaagga tgttcagaat gaaactcatg gaaacactta accagtgcat
     5161 aaacgctggt catgaaatga cgaaggctat cgccattgca cagtttaatg atgacagccc
     5221 ggaagcgagg aaaataaccc ggcgctggag aataggtgaa gcagcggatt tagttggggt
     5281 ttcttctcag gctatcagag atgccgagaa agcagggcga ctaccgcacc cggatatgga
     5341 aattcgagga cgggttgagc aacgtgttgg ttatacaatt gaacaaatta atcatatgcg
     5401 tgatgtgttt ggtacgcgat tgcgacgtgc tgaggacgta tttccaccgg tgatcggggt
     5461 tgctgcccat aaaggtggcg tttacaaaac ctcagtttct gttcatcttg ctcaggatct
     5521 ggctctgaag gggctacgtg ttttgctcgt ggaaggtaac gacccccagg gaacagcctc
     5581 aatgtatcac ggatgggtac cagatcttca tattcatgca gaagatactc tcctgccttt
     5641 ctatcttggg gaaaaggacg atgtcactta tgcaataaag cccacttgct ggccggggct
     5701 tgacattatt ccttcctgtc tggctctgca ccgtattgaa actgagttaa tgggcaaatt
     5761 tgatgaaggt aaactgccca ccgatccaca cctgatgctc cgactggcca ttgaaactgt
     5821 tgctcatgac tatgatgtca tagttattga cagcgcgcct aacctgggta tcggcacgat
     5881 taatgtcgta tgtgctgctg atgtgctgat tgttcccacg cctgctgagt tgtttgacta
     5941 cacctccgca ctgcagtttt tcgatatgct tcgtgatctg ctcaagaacg ttgatcttaa
     6001 agggttcgag cctgatgtac gtattttgct taccaaatac agcaatagta atggctctca
     6061 gtccccgtgg atggaggagc aaattcggga tgcctgggga agcatggttc taaaaaatgt
     6121 tgtacgtgaa acggatgaag ttggtaaagg tcagatccgg atgagaactg tttttgaaca
     6181 ggccattgat caacgcagtt caactggtgc ctggagaaat gctctttcta tttgggaacc
     6241 tgtctgcaat gaaattttcg atcgtctgat taaaccacgc tgggagatta gataatgaag
     6301 cgtgcgcctg ttattccaaa acatacgctc aatactcaac cggttgaaga tacttcgtta
     6361 tcgacaccag ctgccccgat ggtggattcg ttaattgcgc gcgtaggagt aatggctcgc
     6421 ggtaatgcca ttactttgcc tgtatgtggt cgggatgtga agtttactct tgaagtgctc
     6481 cggggtgata gtgttgagaa aacctctcgg gtatggtcag gtaatgaacg tgaccaggag
     6541 ctgcttactg aggacgcact ggatgatctc atcccttctt ttctactgac tggtcaacag
     6601 acaccggcgt tcggtcgaag agtatctggt gtcatagaaa ttgccgatgg gagtcgccgt
     6661 cgtaaagctg ctgcacttac cgaaagtgat tatcgtgttc tggttggcga gctggatgat
     6721 gagcagatgg ctgcattatc cagattgggt aacgattatc gcccaacaag tgcttatgaa
     6781 cgtggtcagc gttatgcaag ccgattgcag aatgaatttg ctggaaatat ttctgcgctg
     6841 gctgatgcgg aaaatatttc acgtaagatt attacccgct gtatcaacac cgccaaattg
     6901 cctaaatcag ttgttgctct tttttctcac cccggtgaac tatctgcccg gtcaggtgat
     6961 gcacttcaaa aagcctttac agataaagag gaattactta agcagcaggc atctaacctt
     7021 catgagcaga aaaaagctgg ggtgatattt gaagctgaag aagttatcac tcttttaact
     7081 tctgtgctta aaacgtcatc tgcatcaaga actagtttaa gctcacgaca tcagtttgct
     7141 cctggagcga cagtattgta taagggcgat aaaatggtgc ttaacctgga caggtcacgt
     7201 gttccaactg agtgtataga gaaaattgag gccattctta aggaacttga aaagccagca
     7261 ccctgatgcg accacgtttt agtctacgtt tatctgtctt tacttaatgt cctttgttac
     7321 aggccagaaa gcataactgg cctgaatatt ctctctgggc ccactgttcc acttgtatcg
     7381 tcggtctgat aatcagactg ggaccacggt cccactcgta tcgtcggtct gattattagt
     7441 ctgggaccac ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacggtccca
     7501 ctcgtatcgt cggtctgata atcagactgg gaccacggtc ccactcgtat cgtcggtctg
     7561 attattagtc tgggaccatg gtcccactcg tatcgtcggt ctgattatta gtctgggacc
     7621 acggtcccac tcgtatcgtc ggtctgatta ttagtctgga accacggtcc cactcgtatc
     7681 gtcggtctga ttattagtct gggaccacgg tcccactcgt atcgtcggtc tgattattag
     7741 tctgggacca cgatcccact cgtgttgtcg gtctgattat cggtctggga ccacggtccc
     7801 acttgtattg tcgatcagac tatcagcgtg agactacgat tccatcaatg cctgtcaagg
     7861 gcaagtattg acatgtcgtc gtaacctgta gaacggagta acctcggtgt gcggttgtat
     7921 gcctgctgtg gattgctgct gtgtcctgct tatccacaac attttgcgca cggttatgtg
     7981 gacaaaatac ctggttaccc aggccgtgcc ggcacgttaa ccgggctgca tccgatgcaa
     8041 gtgtgtcgct gtcgacgcaa gcggccgca
//
