LOCUS       sequence-151863-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to delete part of the mat1-M locus and fix the mating type in 
            natural S. pombe isolates.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5729)
  AUTHORS   Avelar AT, Perfeito L, Gordo I, Ferreira MG
  TITLE     Genome architecture is a selectable trait that can be maintained by 
            antagonistic pleiotropy.
  JOURNAL   Nat Commun. 2013;4:2235. doi: 10.1038/ncomms3235.
  PUBMED    23974178
REFERENCE   2  (bases 1 to 5729)
  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: "Nat 
            Commun."; date: "2013"; volume: "4:2235. doi"; pages: " 
            10.1038/ncomms3235"
COMMENT     Sequence Label: pFA6a-pcr1M-natMX6-pcr2M
FEATURES             Location/Qualifiers
     source          1..5729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1562..2684
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1562..1905
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1906..2481
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVISYSAWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFAGERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGERQALYMSMPCP"
     terminator      2487..2684
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3354..3373)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3355..3373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3460..3477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3631..4219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3711..3730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4390..5181,5182..5250))
                     /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     5013..5032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5251..5355)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5423..5441
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5479..5501)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5601..5620
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5701..5719
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5701..5718
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cagctgaagc tttagacggg aattctttag gggaccccac attaagtggg atgagtgctt
       61 gctttgttgt agtagctatg gtaactattt ctttgtttgg gtaggaacaa agaaagagaa
      121 aggagagaaa atagtgggtt agccgtgaaa ggaaaagaga gtgagaagaa gggtaggaaa
      181 agaagagaga gtagttgaag gaaaaatatt ggaagaggta gtatttttct gtatgcatat
      241 gcatatgcat ttgtatagca tacaaccttt aatttaatag ccatacaacc taatagccag
      301 aaacgccttc ccaatttact gaaccctgct tatatgtagt ttataattgt tgtgtctttt
      361 ttttacgttt aagttgtaat ttgatgttag aatttattac aatggatgca agttttaact
      421 caatttgatc acataggatt tgaattgtaa aaaggtcttc tgctgacatt attagtatgt
      481 attgtgcttt agattaaatt attgtgtttt agtgtatttt gttcctttgt ttttgggttt
      541 ctttgttctg taagtagtta aataccctcc acactctaca cctaccttgc actcacaact
      601 acaatggact cacaccaaga attatcagcg ggttccccta tttcctacga ctttttagat
      661 ccagactggt gttttaaaag ataccttaca aaggatgctc ttcattcaat tgaaactggc
      721 aaaggtgctg cttactttgt tcctgatggc tttactccta tactcattcc taactcacaa
      781 agctacctgt tggatgggaa ttctgcacaa ctgccaagac ctcaaccaat atccttcaca
      841 ttggatcaat gtaaagttcc cggctatatt ttgaagagtt tacggaaaga cactacgtct
      901 actgaacgta ctccgagacc ccctaatgct tttatattat acaggaagga aaaacatgct
      961 acattattaa agtctaaccc ttccatcaat aattcacaag tttccaagct ggttggagag
     1021 atgtggagaa acgaatcaaa ggaagtacgt atgcgatatt ttaagatgtc tgagttttat
     1081 aaagcacaac atcagaaaat gtaccctggt tataagtacc agcctagaaa gaataaagtt
     1141 aaaagataat aatggtaata tttatggtat ttaataaaga attttgaaat tgatttaaat
     1201 atgcttctta gagttacatt cactgaagat tataatgtaa tattttgtgt accccatttg
     1261 cgttgagtta ttctatagta attattgtgt gttctattaa cgatgtattg cgatttatat
     1321 ctgttatgct aacataacgt agttctaagc actgtaatgc catactgttt tagagggtga
     1381 tgcttcctaa aatctcctta cataaagtaa tacatggatt ttactgccct gattctatcg
     1441 aaatatgctg ttttttttat tcgtttttat ttattttcaa taattttttt gtaatataaa
     1501 tgtatagtct ttctccagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag
     1561 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg
     1621 atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat
     1681 acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga
     1741 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag
     1801 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat
     1861 cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg taccactctt
     1921 gacgacacgg cttaccggta ccgcaccagt gtcccggggg acgccgaggc catcgaggca
     1981 ctggatgggt ccttcaccac cgacaccgtc ttccgcgtca ccgccaccgg ggacggcttc
     2041 accctgcggg aggtgccggt ggacccgccc ctgaccaagg tgttccccga cgacgaatcg
     2101 gacgacgaat cggacgacgg ggaggacggc gacccggact cccggacgtt cgtcgcgtac
     2161 ggggacgacg gcgacctggc gggcttcgtg gtcatctcgt actcggcgtg gaaccgccgg
     2221 ctgaccgtcg aggacatcga ggtcgccccg gagcaccggg ggcacggggt cgggcgcgcg
     2281 ttgatggggc tcgcgacgga gttcgccggc gagcggggcg ccgggcacct ctggctggag
     2341 gtcaccaacg tcaacgcacc ggcgatccac gcgtaccggc ggatggggtt caccctctgc
     2401 ggcctggaca ccgccctgta cgacggcacc gcctcggacg gcgagcggca ggcgctctac
     2461 atgagcatgc cctgccccta atcagtactg acaataaaaa gattcttgtt ttcaagaact
     2521 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     2581 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     2641 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     2701 aacgccgcca tccagtttaa acgagctcga attcgccctt agggtttaaa cagatatttg
     2761 cttcgctacg ctacgcactc cctaccataa tatactcact aatattatac tttcactaca
     2821 tctcattact cattcctacc atacacacta tacttcaaca ctttcctcgc tcaaccactc
     2881 catcaaccat cctctttctc cccatacatc ttgaactcct tacaccaata cttcatatcc
     2941 attagccatt ttcctttata aacatatatt aaacctcatc ccaaatgcat ctacccactc
     3001 tttacattca catccctatt cctttaatat cttacgttcc acttcctcaa ccatagagca
     3061 ttctttataa atcactttaa tatgctttac ttccacacac tactgctacc attgccttgc
     3121 acagtactcc ccttactcac acaccttcaa tcacactcct atctccatac aggacccaac
     3181 atgtattaaa cattatcccc ctacaccatt ttccctcatc atccaatatg tttctactat
     3241 tatcacacac cgtctctacc tcccgccaat tttaactgtc tgatatcgca gggcgaattc
     3301 atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata
     3361 gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc
     3421 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     3841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4501 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5221 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5281 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5341 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5401 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5461 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5521 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5581 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga
     5641 catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     5701 atttaggtga cactatagaa cgcggccgc
//
