LOCUS       sequence-247487-        3851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 6 module- NatMX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3851)
  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 3851)
  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: "PLoS One.";
            date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 
            10.1371/journal.pone.0220694. eCollection 2019"
COMMENT     Sequence Label: pMV12
FEATURES             Location/Qualifiers
     source          1..3851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        453..795
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             796..1368
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVISYSAWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFAGERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1374..1571
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1630..1646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1630..1646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1643..1665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1654..1670
                     /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(1678..1684,1685..1702,1703..1708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1723..1744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1861..1878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2032..2620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2112..2131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2791..3582,3583..3651))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGFRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3414..3433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3652..3756)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3824..3842
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tatcggatcc cggtctcgag caacatggag gcccagaata ccctccttga
      481 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
      541 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
      601 aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt
      661 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
      721 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
      781 cgaacataaa caaccatggg taccactctt gacgacacgg cttaccggta ccgcaccagt
      841 gtcccggggg acgccgaggc catcgaggca ctggatgggt ccttcaccac cgacaccgtc
      901 ttccgcgtca ccgccaccgg ggacggcttc accctgcggg aggtgccggt ggacccgccc
      961 ctgaccaagg tgttccccga cgacgaatcg gacgacgaat cggacgacgg ggaggacggc
     1021 gacccggact cccggacgtt cgtcgcgtac ggggacgacg gcgacctggc gggcttcgtg
     1081 gtcatctcgt actcggcgtg gaaccgccgg ctgaccgtcg aggacatcga ggtcgccccg
     1141 gagcaccggg ggcacggggt cgggcgcgcg ttgatggggc tcgcgacgga gttcgccggc
     1201 gagcggggcg ccgggcacct ctggctggag gtcaccaacg tcaacgcacc ggcgatccac
     1261 gcgtaccggc ggatggggtt caccctctgc ggcctggaca ccgccctgta cgacggcacc
     1321 gcctcggacg gcgagcaggc gctctacatg agcatgccct gcccctaatc agtactgaca
     1381 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     1441 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     1501 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     1561 tcgctatact gccatagaga ccgggcccgt cgactgcaga ggcctgcatg caagcttggc
     1621 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1681 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1741 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1801 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1861 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1921 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1981 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2041 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2101 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2161 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2221 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2281 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2341 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2401 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2461 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2521 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2581 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2641 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2701 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2761 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2821 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2881 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gaaacccacg
     2941 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3001 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3061 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3121 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3181 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3241 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3301 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3361 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3421 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3481 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3541 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3601 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3661 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3721 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3781 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3841 gccctttcgt c
//
