LOCUS       sequence-252835-        3921 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Position 6 module- KanMX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3921)
  AUTHORS   Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G
  TITLE     COSPLAY: An expandable toolbox for combinatorial and swift 
            generation of expression plasmids in yeast.
  JOURNAL   PLoS One. 2019 Aug 28;14(8):e0220694. doi: 
            10.1371/journal.pone.0220694. eCollection 2019.
  PUBMED    31461461
REFERENCE   2  (bases 1 to 3921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 
            10.1371/journal.pone.0220694. eCollection 2019"
COMMENT     Sequence Label: pMV18
FEATURES             Location/Qualifiers
     source          1..3921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..380
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             381..1190
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(448..467)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(1249..1265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1249..1265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1262..1284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1273..1289
                     /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)."
     promoter        complement(join(1297..1303,1304..1321,1322..1327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1342..1363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1480..1497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1651..2239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1731..1750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2410..3201,3202..3270))
                     /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     3033..3052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3271..3375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3443..3461
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3499..3521)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3621..3640
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3834..3856
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3848..3865
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3849..3865
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
       61 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
      121 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
      181 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
      241 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
      301 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
      361 acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa
      421 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
      481 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
      541 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
      601 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
      661 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt
      721 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
      781 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
      841 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
      901 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
      961 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     1021 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     1081 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     1141 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa ccatagagac
     1201 cgggcccgtc gactgcagag gcctgcatgc aagcttggcg taatcatggt catagctgtt
     1261 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1321 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1381 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1441 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1501 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1561 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1621 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2161 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2341 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2401 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2461 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2521 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     2581 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2641 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2701 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2761 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2821 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2881 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2941 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3001 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3061 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3121 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3181 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3241 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3301 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3361 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattgt
     3421 tatcacgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     3481 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3541 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3601 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     3661 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     3721 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     3781 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     3841 acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa tgcatctaga
     3901 tatcggatcc cggtctcgag c
//
