LOCUS       sequence-151861-        5873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to delete part of the mat1-P 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 5873)
  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 5873)
  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-pcr1P-natMX6-pcr2P
FEATURES             Location/Qualifiers
     source          1..5873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1696..2818
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1696..2039
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2040..2615
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVISYSAWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFAGERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGERQALYMSMPCP"
     terminator      2621..2818
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3498..3517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3499..3517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3604..3621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3775..4363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3855..3874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4534..5325,5326..5394))
                     /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     5157..5176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5395..5499)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5567..5585
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5623..5645)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5745..5764
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5845..5863
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5845..5862
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cagctgaagc ttgtttattc tatgactaca ggtttgtcaa catgttttat tgcgactgtt
       61 taatttttgt atttgttata ccgatatatt tttaatcagt attatatttt cttaaataac
      121 ttaataacga acttgacaat gaaacgttcc taattctcat tagacgggaa ttctttaggg
      181 gaccccacat taagtgggat gagtgcttgc tttgttgtag tagctatggt aactatttct
      241 ttgtttgggt aggaacaaag aaagagaaag gagagaaaat agtgggttag ccgtgaaagg
      301 aaaagagagt gagaagaagg gtaggaaaag aagagagagt agttgaagga aaaatattgg
      361 aagaggtagt atttttctgt atgcatatgc atatgcattt gtatagcata caacctttaa
      421 tttaatagcc atacaaccta atagccagaa acgccttccc aatttactga accctgctta
      481 tatgtagttt ataattgttg tgtccattta atctttttga atatagtatg cgctctaact
      541 tggcaatttg tctcctaacg tcctgataat gactgagttc agctaattgt gaccaggcaa
      601 gtctaacatt ttgtggtaat gtatgccaaa ttattgagca tttggctgaa gcacctgccc
      661 aatctcctga tagattcaaa attttgtgta gcctacttct aaatagcatg aatccatttt
      721 tgtagattgt ggttttgttt cctttaattt gtttcttctt ttgtaaagtt tctggagtca
      781 aaattggcaa gaaaataggt gctcttaatc ttggatccat tttattattg tgcgtattat
      841 ggcttggtgt atttggagga gtgcaactct gctactgctg tgggcgtttt atatcactaa
      901 tgctttgttc cctctttctt tgttccttat gcgactttat atactctatg gttagtaaca
      961 ccaatacaat ttctgagttt tattttaagc aattcattat gaaacgagta gctgttttat
     1021 taaagactgt aatgtgtgaa tttctcaaat gtgattacaa tggatacgat agaatcattt
     1081 ctcttttgcg gcgtattctt actctaattt gtactccaaa cttaaatggg ttgactatta
     1141 aacgagtaat tgattctatg cagtctttgg aatacattaa acaaacatgt aatttcaaac
     1201 ttcaaatgtg catttccagt atggctttta aaagaaacaa tgctttacag aattgtaatc
     1261 actatgcttg gtgtgatgat cattgttcag atattggaag accgatgact accgtacgcg
     1321 gtcagtgttc caagtgcact aaaccccact tgatgcgttg gcttttgttg cactatgaca
     1381 atccatatcc ctctaactct gagttttacg acctgtcagc ggctactgga ttaacgcgta
     1441 ctcaattacg taactggttt tctaaccgac gacgttgaat tatgttagct tagaattgtt
     1501 agtaatatat ccaattcctt cttgtatatg ttatacatca gttttaatgg gccaattcta
     1561 cgaagtttta aggtattatt cattcggttt ttgtttttca taaatttttt ttttgtaata
     1621 taaatgtata gtctttctcc tttgttttct agatctgttt agcttgcctc gtccccgccg
     1681 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc
     1741 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa
     1801 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg
     1861 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc
     1921 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata
     1981 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca
     2041 tgggtaccac tcttgacgac acggcttacc ggtaccgcac cagtgtcccg ggggacgccg
     2101 aggccatcga ggcactggat gggtccttca ccaccgacac cgtcttccgc gtcaccgcca
     2161 ccggggacgg cttcaccctg cgggaggtgc cggtggaccc gcccctgacc aaggtgttcc
     2221 ccgacgacga atcggacgac gaatcggacg acggggagga cggcgacccg gactcccgga
     2281 cgttcgtcgc gtacggggac gacggcgacc tggcgggctt cgtggtcatc tcgtactcgg
     2341 cgtggaaccg ccggctgacc gtcgaggaca tcgaggtcgc cccggagcac cgggggcacg
     2401 gggtcgggcg cgcgttgatg gggctcgcga cggagttcgc cggcgagcgg ggcgccgggc
     2461 acctctggct ggaggtcacc aacgtcaacg caccggcgat ccacgcgtac cggcggatgg
     2521 ggttcaccct ctgcggcctg gacaccgccc tgtacgacgg caccgcctcg gacggcgagc
     2581 ggcaggcgct ctacatgagc atgccctgcc cctaatcagt actgacaata aaaagattct
     2641 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     2701 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     2761 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     2821 gtcgattcga tactaacgcc gccatccagt ttaaacgagc tcgaattcgc ccttgggttt
     2881 aaacccctct aacgagatat ttgcttcgct acgctacgca ctccctacca taatatactc
     2941 actaatatta tactttcact acatctcatt actcattcct accatacaca ctatacttca
     3001 acactttcct cgctcaacca ctccatcaac catcctcttt ctccccatac atcttgaact
     3061 ccttacacca atacttcata tccattagcc attttccttt ataaacatat attaaacctc
     3121 atcccaaatg catctaccca ctctttacat tcacatccct attcctttaa tatcttacgt
     3181 tccacttcct caaccataga gcattcttta taaatcactt taatatgctt tacttccaca
     3241 cactactgct accattgcct tgcacagtac tccccttact cacacacctt caatcacact
     3301 cctatctcca tacaggaccc aacatgtatt aaacattatc cccctacacc attttccctc
     3361 atcatccaat atgtttctac tattatcaca caccgtctct acctcccgcc aattttaact
     3421 gtctagatat cgcagggcga attcatcgat gatatcagat ccactagtgg cctatgcggc
     3481 cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct
     3541 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3601 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3961 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4201 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4321 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4381 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4441 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4501 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4561 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4621 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4681 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4741 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4801 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4861 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4921 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4981 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5041 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5101 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5161 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5221 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5281 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5341 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5401 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5461 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5521 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5581 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5641 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5701 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5761 tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat
     5821 aaccttatgt atcatacaca tacgatttag gtgacactat agaacgcggc cgc
//
