LOCUS       sequence-263754-        6606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration into fission yeast genome at lys3 locus with bsdMX 
            selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6606)
  AUTHORS   Vjestica A, Marek M, Nkosi PJ, Merlini L, Liu G, Berard M, 
            Billault-Chaumartin I, Martin SG
  TITLE     A toolbox of stable integration vectors in the fission yeast 
            Schizosaccharomyces pombe.
  JOURNAL   J Cell Sci. 2020 Jan 8;133(1). pii: jcs.240754. doi: 
            10.1242/jcs.240754.
  PUBMED    31801797
REFERENCE   2  (bases 1 to 6606)
  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: "J Cell 
            Sci."; date: "2020-01-8"; volume: "133(1). pii: jcs.240754. doi"; 
            pages: " 10.1242/jcs.240754"
COMMENT     Sequence Label: pAV0586
FEATURES             Location/Qualifiers
     source          1..6606
                     /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"
     rep_origin      complement(2526..2981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2663..2684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2875..2894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3107..3129
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3121..3138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3122..3138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3148..3167
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3148..3166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3191..3207
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3192..3208
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3242..3258)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3242..3258)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        3345..3688
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4087..4284
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4346..4366)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4346..4364)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4385..4401)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4385..4401)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4398..4420)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4409..4425
                     /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(4433..4439,4440..4457,4458..4463))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4478..4499
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4616..4633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4787..5375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4867..4886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5546..6337,6338..6406))
                     /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     6169..6188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6407..6511)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6579..6597
                     /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 accataaccg gggccttacg aagttaaatt ataggcacag acatctttgg cattggaaag
      241 atttagtaac aatcataatg tagcacttct taaaaaatct aattttagaa caaaaaggct
      301 taaaaaagaa tggttccgct cttttaaaaa atagtaaaat gtacaatagc aaggttttcg
      361 taggtcgacg gacataacaa ggggaaagag aaataaagca tgtaaacgtc aataaaacat
      421 ctttgattct ggatcaatca tatgccaaat gaagaaaaat agcatttata acaacatgaa
      481 accatatttg agacctttta acgatgaaaa tttattaaat aaaagataaa aaagaaagaa
      541 accgttcaag ttcaatgaaa atttcgtcgt gaagtagaaa ccaagataat ggaaaggctt
      601 cgcaaagcca caaattttcc aatagtcatt ataactgaag attaaggaca gtatactagt
      661 aaaaatcaag tgaattgtgc ctaggatagg tagctaacgt gactgccact gaacctgtgt
      721 agcgtattag atccatcaag atcccttctg ctcatatcat tgctgatctg taaattgacc
      781 gcatcaaatt tgatctcatt tatatcgaga ttgatctata ttattttgcc aatttgcagt
      841 atataaactt atttgattaa gtttttggtc tccttcgcca cgtcttcatt ttgttagttt
      901 aaattttttt gactaactta gaaagggaag tttttaaaaa cgttgaaata ctttttcgat
      961 ttaaaaatat atatttaatt aattttctgt tccaaaatgg ttgctcctca tctttggtta
     1021 cgtgctgaaa ccaaaccttt agaagagcgc agtgctttga ctcctagaac tgccaagata
     1081 ctcgccgatg ctggatttca aattaccatt gagcgctctt cccaacgtgc tttcaaggac
     1141 aaagaatttg agcgccttgg tttccccatg gttcctgagg gctcatggcg acatgcacct
     1201 aaggacgcgt acattattgg tctcaaagag cttccggaaa atgataacag tcctttgaag
     1261 catactcata ttcaatttgc tcattgctac aaaaaccagg aaggttggcg tgaagtcttg
     1321 tcccgattcc ccgctggcaa tggtcttcta tatgatttgg aatttttgca agatgacaat
     1381 ggcagacgtg ttgccgcttt tggttaccac gctggttttg ctggatcagc tatctcatgt
     1441 ttagtttggg cacatcaatt attgcatcct aacaaacaat tccctgctat ccgtccgttc
     1501 cccaacgaaa aatctttggt tcgccatgtt gctcgtcaag tacgtttagc tttgaaaaag
     1561 aataacaacc agtatcctcg tattctggtt attggggctc ttggtcgttg tggtactgga
     1621 gcgtgcgatt tagcatcaaa gattggtatt cccttcgata acattcttcg ctgggacatt
     1681 aatgaaacta agaagggtgg tccctttacc gaaattacgg aatcagacat atttgttaac
     1741 tgcatttatc tgagcatgcc tattcccaaa ttctgtaccg tcgagtcgtt aaatgtacct
     1801 aaccgtaaat tgcgtgttgt ttgtgatgtt agctgtgaca ctaccaatcc taacaatcct
     1861 attccaattt ataatgtaaa cactacattt gatcatccta cagttgaggt taaaggtgtc
     1921 acaactcctc ctcctttaga ggtcatttct attgatcatc ttcctacttt attgccaagg
     1981 gaatcttccg aagcatttag tgaagctcta attcctagtc ttttggcttt gaaggacgtc
     2041 gacaatgctc ccgtttgggt cagagctaaa aaactttatg aaaccatggt tcaaaagctt
     2101 taaaataaat gttatttaat ttcaaagttc cgggtccttt atattgtttc accaaattat
     2161 tgaatattat tttctcaaaa attaacttta gaatacaatt tagacggcat tttcgatgct
     2221 ctggacattt tttccacgta cttgttaaat tgtagcggtg ttgcataata ctaccttttg
     2281 atttgttatt cacgttacaa tatatatttg actaatttaa atatgataat atacacattt
     2341 ctctgcaaac ctcctttata cttttcgccg gtcattgatt aataattaag aaacaggatt
     2401 tcatcctgca tttgccactt acctatatgg taaatataaa tcttgaaact ttccccaata
     2461 tttattgctt gctagggcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2521 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2581 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2641 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2701 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2761 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2821 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2881 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2941 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     3001 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     3061 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     3121 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
     3181 gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg
     3241 ggatccacta gttctagagc ggccgccacc gcggtggtta attaaggcgc gccagatctg
     3301 agctcgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
     3361 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     3421 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     3481 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     3541 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     3601 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     3661 ctcacatcac atccgaacat aaacaaccat gcctttgtct caagaagaat ccaccctcat
     3721 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     3781 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     3841 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     3901 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     3961 tcgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     4021 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     4081 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     4141 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     4201 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     4261 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccacctgca
     4321 ggtaatcacg tgccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     4381 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4441 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4501 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4561 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4621 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4681 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4741 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4801 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4861 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4921 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4981 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5041 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5101 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5161 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5221 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5281 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5341 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5401 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5461 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5521 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5581 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5641 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5701 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5761 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5821 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5881 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5941 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6001 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6061 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6121 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6181 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6241 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6301 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6361 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6421 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6481 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6541 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6601 ttcgtc
//
