LOCUS       sequence-225130-        7978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12392, MetClo assembly vector with F replication origin and 
            chloramphenicol resistance for BsaI-based MetClo, adaptor sequence 
            type a-b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7978)
  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 7978)
  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: pMXBC_aFb
FEATURES             Location/Qualifiers
     source          1..7978
                     /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        1808..1910
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1911..2570
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1977..1996)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3385..3604
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3695..4450
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4453..4703
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5029..6204
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6204..7175
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    7248..7721
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
ORIGIN
        1 cactctgtga cttggtctca aggtggagtg 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 acgcttgcct gagaccaagt cacgaagtgt
     1681 gcggccgcct tcagcggccg cacgaataaa tacctgtgac ggaagatcac ttcgcagaat
     1741 aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat
     1801 gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     1861 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     1921 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     1981 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     2041 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     2101 cccgcctgat gaatgctcat ccggagttcc gtatggcaat gaaagacggt gagctggtga
     2161 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     2221 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     2281 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     2341 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     2401 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     2461 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     2521 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa gagctggagc
     2581 ggcgtaaccg tcgcacagga aggacagaga aagcgcggat ctgggaagtg acggacagaa
     2641 cggtcaggac ctggattggg gaggcggttg ccgccgctgc tgctgacggt gtgacgttct
     2701 ctgttccggt cacaccacat acgttccgcc attcctatgc gatgcacatg ctgtatgccg
     2761 gtataccgct gaaagttctg caaagcctga tgggacataa gtccatcagt tcaacggaag
     2821 tctacacgaa ggtttttgcg ctggatgtgg ctgcccggct ccgggtgcag tttgcgatgc
     2881 cggagtctga tgcggttgcg atgctgaaac aattatcctg agaataaatg ccttggcctt
     2941 tatatggaaa tgtggaactg agtggatatg ctgtttttgt ctgttaaaca gagaagctgg
     3001 ctgttatcca ctgagaagcg aacgaaacag tcgggaaaat ctcccattat cgtagagatc
     3061 cgcattatta atctcaggag cctgtgtagc gtttatagga agtagtgttc tgtcatgatg
     3121 cctgcaagcg gtaacgaaaa cgatttgaat atgccttcag gaacaataga aatcttcgtg
     3181 cggtgttacg ttgaagtgga gcggattatg tcagcaatgg acagaacaac ctaatgaaca
     3241 cagaaccatg atgtggtctg tccttttaca gccagtagtg ctcgccgcag tcgagcgaca
     3301 gggcgaagcc ctcgagtgag cgaggaagca ccagggaaca gcacttatat attctgctta
     3361 cacacgatgc ctgaaaaaac ttcccttggg gttatccact tatccacggg gatattttta
     3421 taattatttt ttttatagtt tttagatctt cttttttaga gcgccttgta ggcctttatc
     3481 catgctggtt ctagagaagg tgttgtgaca aattgccctt tcagtgtgac aaatcaccct
     3541 caaatgacag tcctgtctgt gacaaattgc ccttaaccct gtgacaaatt gccctcagaa
     3601 gaagctgttt tttcacaaag ttatccctgc ttattgactc ttttttattt agtgtgacaa
     3661 tctaaaaact tgtcacactt cacatggatc tgtcatggcg gaaacagcgg ttatcaatca
     3721 caagaaacgt aaaaatagcc cgcgaatcgt ccagtcaaac gacctcactg aggcggcata
     3781 tagtctctcc cgggatcaaa aacgtatgct gtatctgttc gttgaccaga tcagaaaatc
     3841 tgatggcacc ctacaggaac atgacggtat ctgcgagatc catgttgcta aatatgctga
     3901 aatattcgga ttgacctctg cggaagccag taaggatata cggcaggcat tgaagagttt
     3961 cgcggggaag gaagtggttt tttatcgccc tgaagaggat gccggcgatg aaaaaggcta
     4021 tgaatctttt ccttggttta tcaaacgtgc gcacagtcca tccagagggc tttacagtgt
     4081 acatatcaac ccatatctca ttcccttctt tatcgggtta cagaaccggt ttacgcagtt
     4141 tcggcttagt gaaacaaaag aaatcaccaa tccgtatgcc atgcgtttat acgaatccct
     4201 gtgtcagtat cgtaagccgg atggctcagg catcgtgtct ctgaaaatcg actggatcat
     4261 agagcgttac cagctgcctc aaagttacca gcgtatgcct gacttccgcc gccgcttcct
     4321 gcaggtctgt gttaatgaga tcaacagcag aactccaatg cgcctctcat acattgagaa
     4381 aaagaaaggc cgccagacga ctcatatcgt attttccttc cgcgatatca cttccatgac
     4441 gacaggatag tctgagggtt atctgtcaca gatttgaggg tggttcgtca catttgttct
     4501 gacctactga gggtaatttg tcacagtttt gctgtttcct tcagcctgca tggattttct
     4561 catacttttt gaactgtaat ttttaaggaa gccaaatttg agggcagttt gtcacagttg
     4621 atttccttct ctttcccttc gtcatgtgac ctgatatcgg gggttagttc gtcatcattg
     4681 atgagggttg attatcacag tttattactc tgaattggct atccgcgtgt gtacctctac
     4741 ctggagtttt tcccacggtg gatatttctt cttgcgctga gcgtaagagc tatctgacag
     4801 aacagttctt ctttgcttcc tcgccagttc gctcgctatg ctcggttaca cggctgcggc
     4861 gagcgctagt gataataagt gactgaggta tgtggtgttc ttatctcctt ttgtagtgtt
     4921 gctcttattt taaacaactt tgcggttttt tgatgacttt gcgattttgt tgttgctttg
     4981 cagtaaattg caagatttaa taaaaaaacg caaagcaatg attaaaggat gttcagaatg
     5041 aaactcatgg aaacacttaa ccagtgcata aacgctggtc atgaaatgac gaaggctatc
     5101 gccattgcac agtttaatga tgacagcccg gaagcgagga aaataacccg gcgctggaga
     5161 ataggtgaag cagcggattt agttggggtt tcttctcagg ctatcagaga tgccgagaaa
     5221 gcagggcgac taccgcaccc ggatatggaa attcgaggac gggttgagca acgtgttggt
     5281 tatacaattg aacaaattaa tcatatgcgt gatgtgtttg gtacgcgatt gcgacgtgct
     5341 gaggacgtat ttccaccggt gatcggggtt gctgcccata aaggtggcgt ttacaaaacc
     5401 tcagtttctg ttcatcttgc tcaggatctg gctctgaagg ggctacgtgt tttgctcgtg
     5461 gaaggtaacg acccccaggg aacagcctca atgtatcacg gatgggtacc agatcttcat
     5521 attcatgcag aagatactct cctgcctttc tatcttgggg aaaaggacga tgtcacttat
     5581 gcaataaagc ccacttgctg gccggggctt gacattattc cttcctgtct ggctctgcac
     5641 cgtattgaaa ctgagttaat gggcaaattt gatgaaggta aactgcccac cgatccacac
     5701 ctgatgctcc gactggccat tgaaactgtt gctcatgact atgatgtcat agttattgac
     5761 agcgcgccta acctgggtat cggcacgatt aatgtcgtat gtgctgctga tgtgctgatt
     5821 gttcccacgc ctgctgagtt gtttgactac acctccgcac tgcagttttt cgatatgctt
     5881 cgtgatctgc tcaagaacgt tgatcttaaa gggttcgagc ctgatgtacg tattttgctt
     5941 accaaataca gcaatagtaa tggctctcag tccccgtgga tggaggagca aattcgggat
     6001 gcctggggaa gcatggttct aaaaaatgtt gtacgtgaaa cggatgaagt tggtaaaggt
     6061 cagatccgga tgagaactgt ttttgaacag gccattgatc aacgcagttc aactggtgcc
     6121 tggagaaatg ctctttctat ttgggaacct gtctgcaatg aaattttcga tcgtctgatt
     6181 aaaccacgct gggagattag ataatgaagc gtgcgcctgt tattccaaaa catacgctca
     6241 atactcaacc ggttgaagat acttcgttat cgacaccagc tgccccgatg gtggattcgt
     6301 taattgcgcg cgtaggagta atggctcgcg gtaatgccat tactttgcct gtatgtggtc
     6361 gggatgtgaa gtttactctt gaagtgctcc ggggtgatag tgttgagaaa acctctcggg
     6421 tatggtcagg taatgaacgt gaccaggagc tgcttactga ggacgcactg gatgatctca
     6481 tcccttcttt tctactgact ggtcaacaga caccggcgtt cggtcgaaga gtatctggtg
     6541 tcatagaaat tgccgatggg agtcgccgtc gtaaagctgc tgcacttacc gaaagtgatt
     6601 atcgtgttct ggttggcgag ctggatgatg agcagatggc tgcattatcc agattgggta
     6661 acgattatcg cccaacaagt gcttatgaac gtggtcagcg ttatgcaagc cgattgcaga
     6721 atgaatttgc tggaaatatt tctgcgctgg ctgatgcgga aaatatttca cgtaagatta
     6781 ttacccgctg tatcaacacc gccaaattgc ctaaatcagt tgttgctctt ttttctcacc
     6841 ccggtgaact atctgcccgg tcaggtgatg cacttcaaaa agcctttaca gataaagagg
     6901 aattacttaa gcagcaggca tctaaccttc atgagcagaa aaaagctggg gtgatatttg
     6961 aagctgaaga agttatcact cttttaactt ctgtgcttaa aacgtcatct gcatcaagaa
     7021 ctagtttaag ctcacgacat cagtttgctc ctggagcgac agtattgtat aagggcgata
     7081 aaatggtgct taacctggac aggtcacgtg ttccaactga gtgtatagag aaaattgagg
     7141 ccattcttaa ggaacttgaa aagccagcac cctgatgcga ccacgtttta gtctacgttt
     7201 atctgtcttt acttaatgtc ctttgttaca ggccagaaag cataactggc ctgaatattc
     7261 tctctgggcc cactgttcca cttgtatcgt cggtctgata atcagactgg gaccacggtc
     7321 ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg tatcgtcggt
     7381 ctgattatta gtctgggacc acggtcccac tcgtatcgtc ggtctgataa tcagactggg
     7441 accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccatgg tcccactcgt
     7501 atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg gtctgattat
     7561 tagtctggaa ccacggtccc actcgtatcg tcggtctgat tattagtctg ggaccacggt
     7621 cccactcgta tcgtcggtct gattattagt ctgggaccac gatcccactc gtgttgtcgg
     7681 tctgattatc ggtctgggac cacggtccca cttgtattgt cgatcagact atcagcgtga
     7741 gactacgatt ccatcaatgc ctgtcaaggg caagtattga catgtcgtcg taacctgtag
     7801 aacggagtaa cctcggtgtg cggttgtatg cctgctgtgg attgctgctg tgtcctgctt
     7861 atccacaaca ttttgcgcac ggttatgtgg acaaaatacc tggttaccca ggccgtgccg
     7921 gcacgttaac cgggctgcat ccgatgcaag tgtgtcgctg tcgacgcaag cggccgca
//
