LOCUS       sequence-224790-        2432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12356, MetClo assembly vector with p15A 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 2432)
  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 2432)
  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: pMXLC_aFb
FEATURES             Location/Qualifiers
     source          1..2432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    111..132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(147..152,153..170,171..177)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    185..201
                     /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     190..212
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     209..225
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     209..225
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             221..544
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    238..294
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(295..312)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(295..311)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(304..326)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(520..539)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        819..921
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             922..1581
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(988..1007)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1607..2152
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2053..2072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2269..2286
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtga cttggtctca aggtggagtg agaccggcct agcaggtgtg agtttgcagc
       61 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
      121 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      181 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
      241 cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga attcactggc
      301 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
      361 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      421 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
      481 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
      541 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      601 gctcccggca tccgcttaca gacaagctgt gacaagtacc acctggaacg ccaaaggtct
      661 cacgcttgcc tgagaccaag tcacgaagtg tgcggccgcc ttcagcggcc gcacgaataa
      721 atacctgtga cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc
      781 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc
      841 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
      901 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
      961 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     1021 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     1081 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggagttc
     1141 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     1201 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     1261 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     1321 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     1381 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     1441 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     1501 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     1561 gagtggcagg gcggggcgta agagcctgtt taataagatg atcttcttga gatcgttttg
     1621 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
     1681 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
     1741 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
     1801 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
     1861 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
     1921 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
     1981 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
     2041 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
     2101 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
     2161 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg
     2221 ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg
     2281 agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt
     2341 tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca
     2401 gtatacactc cgctagcgcg caagcggccg ca
//
