LOCUS       sequence-263766-        5555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration into fission yeast genome at lys3 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5555)
  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 5555)
  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: pAV0357
FEATURES             Location/Qualifiers
     source          1..5555
                     /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"
     CDS             complement(2958..3338)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     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"
     misc_feature    3175..3282
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     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"
     primer_bind     complement(3295..3315)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3295..3313)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3334..3350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3334..3350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3347..3369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3358..3374
                     /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(3382..3388,3389..3406,3407..3412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3427..3448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3565..3582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3736..4324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3816..3835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4495..5286,5287..5355))
                     /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     5118..5137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5356..5460)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5528..5546
                     /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 gcggtggagc tccagctttt gttcccttta
     3301 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5461 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5521 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
