LOCUS       sequence-263767-        5726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration into fission yeast genome at ade6 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  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 5726)
  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: pAV0356
FEATURES             Location/Qualifiers
     source          1..5726
                     /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"
     CDS             complement(3129..3509)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     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"
     misc_feature    3346..3453
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     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"
     primer_bind     complement(3466..3486)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3466..3484)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3505..3521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3505..3521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3518..3540)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3529..3545
                     /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(3553..3559,3560..3577,3578..3583))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3598..3619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3736..3753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3907..4495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3987..4006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4666..5457,5458..5526))
                     /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     5289..5308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5527..5631)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5699..5717
                     /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 cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     3481 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3541 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3601 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3661 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3721 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3781 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3841 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4321 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5521 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5581 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5641 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5701 taggcgtatc acgaggccct ttcgtc
//
