LOCUS Exported 2519 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION POC12379, MetClo assembly vector with p15A replication origin and ampicillin resistance for BsaI-based MetClo, adaptor sequence type p-a. ACCESSION . VERSION . KEYWORDS pMXLP_pFa 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2519 /organism="synthetic DNA construct" /mol_type="other DNA" 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 143..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 703..807 /gene="bla" /label=AmpR promoter CDS 808..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 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 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 //