LOCUS       sequence-225080-        2519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  POC12389, MetClo assembly vector with p15A replication origin and 
            ampicillin resistance for BsaI-based MetClo, adaptor sequence type 
            d-q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2519)
  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 2519)
  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_dFq
FEATURES             Location/Qualifiers
     source          1..2519
                     /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             join(808..876,877..1668)
                     /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(1026..1045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1694..2239
                     /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     2140..2159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2356..2373
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtga cttggtctca acgaggagtg 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 ttgataaatg
      781 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      841 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      901 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      961 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1021 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1081 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1141 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1201 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1261 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1321 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1381 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1441 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1501 aaatctggag ccggtgagcg tggttctcgc ggtatcattg cagcactggg gccagatggt
     1561 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1621 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaaga gcctgtttaa
     1681 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     1741 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     1801 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     1861 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     1921 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1981 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     2041 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     2101 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     2161 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     2221 ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc
     2281 ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt
     2341 cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata
     2401 ttctgctgac gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac
     2461 cctcatcagt gccaacatag taagccagta tacactccgc tagcgcgcaa gcggccgca
//
