LOCUS       sequence-224787-        2428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12355, MetClo assembly vector with p15A replication origin and 
            chloramphenicol 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 2428)
  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 2428)
  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_pFa
FEATURES             Location/Qualifiers
     source          1..2428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             217..540
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    234..290
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(291..308)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(291..307)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(300..322)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(516..535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        815..917
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             918..1577
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(984..1003)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1603..2148
                     /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     2049..2068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2265..2282
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtga cttggtctca ggagtgagac cggcctagca ggtgtgagtt tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc
      301 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      361 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      421 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
      481 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
      541 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
      601 ccggcatccg cttacagaca agctgtgaca agtaccacct ggaacgccaa aggtctcacg
      661 ctaggttgag accaagtcac gaagtgtgcg gccgccttca gcggccgcac gaataaatac
      721 ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga
      781 agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
      841 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
      901 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
      961 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     1021 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     1081 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gagttccgta
     1141 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     1201 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     1261 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     1321 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     1381 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     1441 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     1501 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     1561 ggcagggcgg ggcgtaagag cctgtttaat aagatgatct tcttgagatc gttttggtct
     1621 gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt
     1681 tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa
     1741 acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc
     1801 aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac
     1861 gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca
     1921 gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc
     1981 cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag
     2041 ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt
     2101 gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc
     2161 gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc
     2221 gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg
     2281 aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc
     2341 tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat
     2401 acactccgct agcgcgcaag cggccgca
//
