LOCUS       sequence-225039-        2453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12374, MetClo assembly vector with p15A replication origin and 
            kanamycin resistance for BsaI-based MetClo, adaptor sequence type 
            a-q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2453)
  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 2453)
  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: pMXLK_aFq
FEATURES             Location/Qualifiers
     source          1..2453
                     /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        703..807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             808..1602
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1628..2173
                     /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     2074..2093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2290..2307
                     /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 cacgcttgag accaagtcac gaagtgtgcg gccgccttcg tgcgcggaac ccctatttgt
      721 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      781 cttcaataat attgaaaaag gaagagtatg gctaaaatga gaatatcacc ggaattgaaa
      841 aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta
      901 tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa
      961 gggaccacct atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg
     1021 cctgttccaa aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt
     1081 gaggccgatg gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt
     1141 atcgagctgt atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc
     1201 tatacgaata gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg
     1261 gccgatgtgg attgcgaaaa ctgggaagag gacactccat ttaaagatcc gcgcgagctg
     1321 tatgattttt taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg
     1381 ggagacagca acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga
     1441 agcggcaggg cggacaagtg gtatgacatt gccttctgcg tccggtcgct cagggaggat
     1501 atcggggaag aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg
     1561 gagaaaataa aatattatat tttactggat gaattgtttt aggagcctgt ttaataagat
     1621 gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg
     1681 ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact
     1741 ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg
     1801 acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca
     1861 tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga
     1921 acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc
     1981 aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc
     2041 aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc
     2101 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     2161 gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg
     2221 gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg
     2281 accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc
     2341 tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg acaccctcat
     2401 cagtgccaac atagtaagcc agtatacact ccgctagcgc gcaagcggcc gca
//
