LOCUS       sequence-225050-        2523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12380, MetClo assembly vector with p15A replication origin and 
            ampicillin 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 2523)
  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 2523)
  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: pMXLP_aFb
FEATURES             Location/Qualifiers
     source          1..2523
                     /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        707..811
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(812..880,881..1672)
                     /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(1030..1049)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1698..2243
                     /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     2144..2163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /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 ttcgtgcgcg gaacccctat
      721 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccttgata
      781 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
      841 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
      901 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
      961 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1021 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1081 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1141 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1201 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1261 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1321 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1381 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1441 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1501 tgataaatct ggagccggtg agcgtggttc tcgcggtatc attgcagcac tggggccaga
     1561 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     1621 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aagagcctgt
     1681 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
     1741 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
     1801 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     1861 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     1921 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1981 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     2041 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     2101 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     2161 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     2221 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag
     2281 tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg
     2341 cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca
     2401 catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg
     2461 acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc gcaagcggcc
     2521 gca
//
