LOCUS       sequence-263796-        7413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration into fission yeast genome at ade6 locus with hphMX 
            selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7413)
  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 7413)
  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: pAV0585
FEATURES             Location/Qualifiers
     source          1..7413
                     /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(2697..3152)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3046..3065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3278..3300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3292..3309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3293..3309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3319..3338
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3319..3337
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3362..3378
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3363..3379
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3413..3429)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3413..3429)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     gene            3516..5091
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        3516..3859
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3860..4888
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMRLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVGASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      4894..5091
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5153..5173)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5153..5171)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5192..5208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5192..5208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5205..5227)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5216..5232
                     /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(5240..5246,5247..5264,5265..5270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5285..5306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5423..5440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5594..6182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5674..5693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6353..7144,7145..7213))
                     /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     6976..6995
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7214..7318)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7386..7404
                     /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 ggagaatcct gaataatgtg ctgtgaagca gttgaaagaa aattaaagaa
      241 cccaatataa tatgctataa agcaatcaaa aaataatagt aagaaaccca aataaaaccc
      301 cgttgaccat gttcccaatt aaattaacgc atattaatgc aaaaaatgca atattaagac
      361 tagttattaa accgataatt agtcttcaat tattaaaaga tgtagttgat ttgtattatc
      421 tttcaattgc taattctaaa tatattggag aattcaatga gtttatgcat tgaagtttaa
      481 gataacattg gcgctgattg cagcaaaagt aatcacgcat atagctgtag gaccaaaacg
      541 atcatcatat cggaccgcat tatgtttaaa cgttgcttta tatatggata acgacatata
      601 catttatttc ttgaaaaaat agcaaagcta tattgatttt agcgtatgct gctttaaagt
      661 tgcatttcac aatgcttgga aatgtaacga tgacagaatt aatacgatgc aaaactcaaa
      721 taataaactg cgcactaact cactacaata aacaactttg aaaaaaagat tcgttttttc
      781 aacatttacc atctcattaa gctgagctgc caaggtatat acatacttca tcgaatatga
      841 gcgaaaaaca ggttgtaggg atccttggag gtggtcaatt gggccgaatg atggtagagg
      901 cagcccatcg cttaaacatc aaatgcatca tcttggatgc agcaaattct cctgccaaac
      961 aaattgatgg aggacgtgag cacattgatg catcatttac tgaccccgat gcaattgttg
     1021 aactgtctaa gaagtgcacg ttattaacaa ctgaaattga gcatattaac actgatgcct
     1081 tggcagccgt tacgaaatct gttgctgttg aaccctctcc tgcaactctg cgatgcattc
     1141 aagacaaata tcttcaaaaa cagcatttac aggtttttaa gattgcactt cctgaatttt
     1201 gcgatgcacc tgaccaggaa agtgttgaaa aagcaggcca agagtttggt tatccttttg
     1261 tactgaaaag taaaacattg gcttacgacg gtcgtggaaa ttacgttgtt catcaaccat
     1321 ctgagattcc tactgccatc aaagcacttg gtgatcgtcc gctttatgtt gaaaagttcg
     1381 ttcctttctc catggaaatt gcagtgatgg tagtacgcag tttagacgga aaagtttatg
     1441 cttatcctac aactgagacc attcaaaagg ataatgtttg tcatttagta tatgcccctg
     1501 ctcgtcttcc cttctcaatt caacagcgtg ctcaaaccct tgccatggat gcggtgcgca
     1561 cttttgaagg cgctggtata tatggtgtag agatgtttgt tttgcaagat ggtgagacca
     1621 ttttactcaa cgaaattgct cctcggcctc acaattcagg tcactacacc attgaagctt
     1681 gcccaacttc tcagtttgaa gctcacttac gggccatatg tggtcttcct ttcagcgaaa
     1741 tcaacaccca actctcgact tccacaactc atgcgttgat ggtaaatatt ttaggtactg
     1801 atgatcctga ttatgtttca aagatcgcta aacgttctct gtccattccc ggtgcaactt
     1861 tgcatcttta tggtaaagct gaatctagaa agggtcgcaa gatgggacac gttaccatca
     1921 tttctgattc acctcaagaa tgtgaacgta ggtatcagat gcttcttgac gtcaaagatc
     1981 ctgtcgaatc acctgttgtt ggtattatca tgggttcgga ttctgattta agcaagatga
     2041 aagatgctgc cgtcatttta gatgaattca aggtgcctta cgaacttact attgtttcag
     2101 ctcaccgcac accagatcgc atggttactt atgctcgtac cgcagcttca agagggttgc
     2161 gtgtgattat tgctggtgct ggtggtgccg ctcatttgcc tggtatggtt gctgcaatga
     2221 cacctcttcc agtaatcggc gttcctgtaa aaggaagcac tcttgacgga gttgactctc
     2281 ttcactctat tgttcagatg cctcgaggtg tccctgtcgc cactgttgct atcaataata
     2341 gccaaaatgc cggtatttta gcctgtcgta tacttgctac atttcaaccc tcccttttgg
     2401 ctgctatgga gagctttatg gacaatatga aagagattgt tttagaaaag gctgataaat
     2461 tagaaaaagt tggttggaaa aattattctg cataggcgac catagacata actgttaaat
     2521 gattttgctt gctttttgtt ttaaataata ttttattttg aaattctaga ttgtaaaatg
     2581 tattctaaaa agtgattaaa aattaaaaat aaaaaccccc cgaataatgt gctgcgacgc
     2641 agctagggcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     2701 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2761 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2821 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2881 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2941 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3001 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3061 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3121 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     3181 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3301 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     3361 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact
     3421 agttctagag cggccgccac cgcggtggtt aattaaggcg cgccagatct gagctcgttt
     3481 agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc
     3541 ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc
     3601 ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa
     3661 gcaaaaatta cggctcctcg ctgcggacct gcgagcaggg aaacgctccc ctcacagacg
     3721 cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac
     3781 tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca
     3841 catccgaaca taaacaacca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     3901 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cggctctcgg agggcgaaga
     3961 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     4021 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     4081 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     4141 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4201 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     4261 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4321 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcggtgcgtc
     4381 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     4441 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     4501 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     4561 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     4621 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     4681 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     4741 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     4801 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     4861 cagcactcgt ccgagggcaa aggaataatc agtactgaca ataaaaagat tcttgttttc
     4921 aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt
     4981 agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg
     5041 cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt
     5101 cgatactaac gccgccatcc acctgcaggt aatcacgtgc cagcttttgt tccctttagt
     5161 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5221 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5281 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5401 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5461 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5521 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5581 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5641 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5701 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5761 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5821 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5881 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5941 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6001 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6061 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6121 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6181 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6241 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6301 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6361 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6421 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6481 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6541 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6601 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6661 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6721 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6781 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6841 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6901 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6961 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7021 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7081 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7141 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7201 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7261 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7321 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     7381 ataaaaatag gcgtatcacg aggccctttc gtc
//
