LOCUS       sequence-225169-        8065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12403, MetClo assembly vector with F replication origin and 
            ampicillin resistance for BsaI-based MetClo, adaptor sequence type 
            p-a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8065)
  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 8065)
  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_pFa
FEATURES             Location/Qualifiers
     source          1..8065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      220..808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     709..728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     962..979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1096..1117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1132..1137,1138..1155,1156..1162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1170..1186
                     /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     1175..1197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1194..1210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1194..1210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1206..1529
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    1223..1279
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1280..1297)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1280..1296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1289..1311)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1505..1524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1692..1796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1797..1865,1866..2657)
                     /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(2015..2034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3472..3691
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3782..4537
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4540..4790
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5116..6291
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6291..7262
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    7335..7808
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
ORIGIN
        1 cactctgtga cttggtctca ggagtgagac cggcctagca ggtgtgagtt tgtcagacca
       61 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
      121 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
      181 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      241 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      301 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      361 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      421 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      481 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      541 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      601 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      661 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      721 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      781 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      841 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      901 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      961 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     1021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1081 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     1141 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     1201 cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc
     1261 gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1321 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1381 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     1441 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     1501 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     1561 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgacaa
     1621 gtaccacctg gaacgccaaa ggtctcacgc taggttgaga ccaagtcacg aagtgtgcgg
     1681 ccgccttcgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1741 cgctcatgag acaataacct tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1801 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1861 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1921 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1981 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2041 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2101 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2161 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2221 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2281 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2341 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2401 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2461 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggttctcgcg
     2521 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2581 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2641 tgattaagca ttggtaagag ctggagcggc gtaaccgtcg cacaggaagg acagagaaag
     2701 cgcggatctg ggaagtgacg gacagaacgg tcaggacctg gattggggag gcggttgccg
     2761 ccgctgctgc tgacggtgtg acgttctctg ttccggtcac accacatacg ttccgccatt
     2821 cctatgcgat gcacatgctg tatgccggta taccgctgaa agttctgcaa agcctgatgg
     2881 gacataagtc catcagttca acggaagtct acacgaaggt ttttgcgctg gatgtggctg
     2941 cccggctccg ggtgcagttt gcgatgccgg agtctgatgc ggttgcgatg ctgaaacaat
     3001 tatcctgaga ataaatgcct tggcctttat atggaaatgt ggaactgagt ggatatgctg
     3061 tttttgtctg ttaaacagag aagctggctg ttatccactg agaagcgaac gaaacagtcg
     3121 ggaaaatctc ccattatcgt agagatccgc attattaatc tcaggagcct gtgtagcgtt
     3181 tataggaagt agtgttctgt catgatgcct gcaagcggta acgaaaacga tttgaatatg
     3241 ccttcaggaa caatagaaat cttcgtgcgg tgttacgttg aagtggagcg gattatgtca
     3301 gcaatggaca gaacaaccta atgaacacag aaccatgatg tggtctgtcc ttttacagcc
     3361 agtagtgctc gccgcagtcg agcgacaggg cgaagccctc gagtgagcga ggaagcacca
     3421 gggaacagca cttatatatt ctgcttacac acgatgcctg aaaaaacttc ccttggggtt
     3481 atccacttat ccacggggat atttttataa ttattttttt tatagttttt agatcttctt
     3541 ttttagagcg ccttgtaggc ctttatccat gctggttcta gagaaggtgt tgtgacaaat
     3601 tgccctttca gtgtgacaaa tcaccctcaa atgacagtcc tgtctgtgac aaattgccct
     3661 taaccctgtg acaaattgcc ctcagaagaa gctgtttttt cacaaagtta tccctgctta
     3721 ttgactcttt tttatttagt gtgacaatct aaaaacttgt cacacttcac atggatctgt
     3781 catggcggaa acagcggtta tcaatcacaa gaaacgtaaa aatagcccgc gaatcgtcca
     3841 gtcaaacgac ctcactgagg cggcatatag tctctcccgg gatcaaaaac gtatgctgta
     3901 tctgttcgtt gaccagatca gaaaatctga tggcacccta caggaacatg acggtatctg
     3961 cgagatccat gttgctaaat atgctgaaat attcggattg acctctgcgg aagccagtaa
     4021 ggatatacgg caggcattga agagtttcgc ggggaaggaa gtggtttttt atcgccctga
     4081 agaggatgcc ggcgatgaaa aaggctatga atcttttcct tggtttatca aacgtgcgca
     4141 cagtccatcc agagggcttt acagtgtaca tatcaaccca tatctcattc ccttctttat
     4201 cgggttacag aaccggttta cgcagtttcg gcttagtgaa acaaaagaaa tcaccaatcc
     4261 gtatgccatg cgtttatacg aatccctgtg tcagtatcgt aagccggatg gctcaggcat
     4321 cgtgtctctg aaaatcgact ggatcataga gcgttaccag ctgcctcaaa gttaccagcg
     4381 tatgcctgac ttccgccgcc gcttcctgca ggtctgtgtt aatgagatca acagcagaac
     4441 tccaatgcgc ctctcataca ttgagaaaaa gaaaggccgc cagacgactc atatcgtatt
     4501 ttccttccgc gatatcactt ccatgacgac aggatagtct gagggttatc tgtcacagat
     4561 ttgagggtgg ttcgtcacat ttgttctgac ctactgaggg taatttgtca cagttttgct
     4621 gtttccttca gcctgcatgg attttctcat actttttgaa ctgtaatttt taaggaagcc
     4681 aaatttgagg gcagtttgtc acagttgatt tccttctctt tcccttcgtc atgtgacctg
     4741 atatcggggg ttagttcgtc atcattgatg agggttgatt atcacagttt attactctga
     4801 attggctatc cgcgtgtgta cctctacctg gagtttttcc cacggtggat atttcttctt
     4861 gcgctgagcg taagagctat ctgacagaac agttcttctt tgcttcctcg ccagttcgct
     4921 cgctatgctc ggttacacgg ctgcggcgag cgctagtgat aataagtgac tgaggtatgt
     4981 ggtgttctta tctccttttg tagtgttgct cttattttaa acaactttgc ggttttttga
     5041 tgactttgcg attttgttgt tgctttgcag taaattgcaa gatttaataa aaaaacgcaa
     5101 agcaatgatt aaaggatgtt cagaatgaaa ctcatggaaa cacttaacca gtgcataaac
     5161 gctggtcatg aaatgacgaa ggctatcgcc attgcacagt ttaatgatga cagcccggaa
     5221 gcgaggaaaa taacccggcg ctggagaata ggtgaagcag cggatttagt tggggtttct
     5281 tctcaggcta tcagagatgc cgagaaagca gggcgactac cgcacccgga tatggaaatt
     5341 cgaggacggg ttgagcaacg tgttggttat acaattgaac aaattaatca tatgcgtgat
     5401 gtgtttggta cgcgattgcg acgtgctgag gacgtatttc caccggtgat cggggttgct
     5461 gcccataaag gtggcgttta caaaacctca gtttctgttc atcttgctca ggatctggct
     5521 ctgaaggggc tacgtgtttt gctcgtggaa ggtaacgacc cccagggaac agcctcaatg
     5581 tatcacggat gggtaccaga tcttcatatt catgcagaag atactctcct gcctttctat
     5641 cttggggaaa aggacgatgt cacttatgca ataaagccca cttgctggcc ggggcttgac
     5701 attattcctt cctgtctggc tctgcaccgt attgaaactg agttaatggg caaatttgat
     5761 gaaggtaaac tgcccaccga tccacacctg atgctccgac tggccattga aactgttgct
     5821 catgactatg atgtcatagt tattgacagc gcgcctaacc tgggtatcgg cacgattaat
     5881 gtcgtatgtg ctgctgatgt gctgattgtt cccacgcctg ctgagttgtt tgactacacc
     5941 tccgcactgc agtttttcga tatgcttcgt gatctgctca agaacgttga tcttaaaggg
     6001 ttcgagcctg atgtacgtat tttgcttacc aaatacagca atagtaatgg ctctcagtcc
     6061 ccgtggatgg aggagcaaat tcgggatgcc tggggaagca tggttctaaa aaatgttgta
     6121 cgtgaaacgg atgaagttgg taaaggtcag atccggatga gaactgtttt tgaacaggcc
     6181 attgatcaac gcagttcaac tggtgcctgg agaaatgctc tttctatttg ggaacctgtc
     6241 tgcaatgaaa ttttcgatcg tctgattaaa ccacgctggg agattagata atgaagcgtg
     6301 cgcctgttat tccaaaacat acgctcaata ctcaaccggt tgaagatact tcgttatcga
     6361 caccagctgc cccgatggtg gattcgttaa ttgcgcgcgt aggagtaatg gctcgcggta
     6421 atgccattac tttgcctgta tgtggtcggg atgtgaagtt tactcttgaa gtgctccggg
     6481 gtgatagtgt tgagaaaacc tctcgggtat ggtcaggtaa tgaacgtgac caggagctgc
     6541 ttactgagga cgcactggat gatctcatcc cttcttttct actgactggt caacagacac
     6601 cggcgttcgg tcgaagagta tctggtgtca tagaaattgc cgatgggagt cgccgtcgta
     6661 aagctgctgc acttaccgaa agtgattatc gtgttctggt tggcgagctg gatgatgagc
     6721 agatggctgc attatccaga ttgggtaacg attatcgccc aacaagtgct tatgaacgtg
     6781 gtcagcgtta tgcaagccga ttgcagaatg aatttgctgg aaatatttct gcgctggctg
     6841 atgcggaaaa tatttcacgt aagattatta cccgctgtat caacaccgcc aaattgccta
     6901 aatcagttgt tgctcttttt tctcaccccg gtgaactatc tgcccggtca ggtgatgcac
     6961 ttcaaaaagc ctttacagat aaagaggaat tacttaagca gcaggcatct aaccttcatg
     7021 agcagaaaaa agctggggtg atatttgaag ctgaagaagt tatcactctt ttaacttctg
     7081 tgcttaaaac gtcatctgca tcaagaacta gtttaagctc acgacatcag tttgctcctg
     7141 gagcgacagt attgtataag ggcgataaaa tggtgcttaa cctggacagg tcacgtgttc
     7201 caactgagtg tatagagaaa attgaggcca ttcttaagga acttgaaaag ccagcaccct
     7261 gatgcgacca cgttttagtc tacgtttatc tgtctttact taatgtcctt tgttacaggc
     7321 cagaaagcat aactggcctg aatattctct ctgggcccac tgttccactt gtatcgtcgg
     7381 tctgataatc agactgggac cacggtccca ctcgtatcgt cggtctgatt attagtctgg
     7441 gaccacggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg
     7501 tatcgtcggt ctgataatca gactgggacc acggtcccac tcgtatcgtc ggtctgatta
     7561 ttagtctggg accatggtcc cactcgtatc gtcggtctga ttattagtct gggaccacgg
     7621 tcccactcgt atcgtcggtc tgattattag tctggaacca cggtcccact cgtatcgtcg
     7681 gtctgattat tagtctggga ccacggtccc actcgtatcg tcggtctgat tattagtctg
     7741 ggaccacgat cccactcgtg ttgtcggtct gattatcggt ctgggaccac ggtcccactt
     7801 gtattgtcga tcagactatc agcgtgagac tacgattcca tcaatgcctg tcaagggcaa
     7861 gtattgacat gtcgtcgtaa cctgtagaac ggagtaacct cggtgtgcgg ttgtatgcct
     7921 gctgtggatt gctgctgtgt cctgcttatc cacaacattt tgcgcacggt tatgtggaca
     7981 aaatacctgg ttacccaggc cgtgccggca cgttaaccgg gctgcatccg atgcaagtgt
     8041 gtcgctgtcg acgcaagcgg ccgca
//
