LOCUS       sequence-263762-        7495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in fission yeast from promoter pact1, integrated at ade6 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7495)
  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 7495)
  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: pAV0661
FEATURES             Location/Qualifiers
     source          1..7495
                     /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"
     terminator      4961..5210
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4977..4995)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5235..5255)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5235..5253)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5274..5290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5274..5290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5287..5309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5298..5314
                     /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(5322..5328,5329..5346,5347..5352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5367..5388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5505..5522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5676..6264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5756..5775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6435..7226,7227..7295))
                     /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     7058..7077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7296..7400)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7468..7486
                     /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 cgggcccatt
     3361 taaatgatct acgataatga gacggtgttt gggttgtttt tactgaattt ttcttttatt
     3421 ttatgacatc ttaattttta tgttaatgat tattcttata cttttatcta tctttgatta
     3481 aaactaatat tcattctttt gatcactaat tatttttata gtatacgatt caacccaact
     3541 ataatacatt gtatcaagag aaatttttaa ttagatgtaa tttaatagcc atttgataaa
     3601 aaaaaattac cttttttttt gctagtgtta aattattgat cataacaatg tcgaatgaat
     3661 actaataaaa tagggacacg cgagttgcaa atacttgaaa tacctttgaa gatttgtctg
     3721 tgaattttga taagattttt atgaccatct cttttatatt cgatatcgtg aactttcatt
     3781 ttcttttgtt ttttgctttt caagttaaat aacgcggaaa ttatatttcc ttatatgttt
     3841 ataagagctt ttaaaggatt tgctatatcg cttacagtac tacaaccggt tgctcaatgt
     3901 tatccgtttc cgcggtcttc ttccgtgcgc attctgccgt gaagtgctaa cgctgtgtgt
     3961 ggctttatca aaaaagtggc tcaaatatag caatttcata catatttaaa cacccattat
     4021 acttaaatgt atattttaaa atatataaga gattttcact gtataactgt gtatcttatc
     4081 tatatgtaat caacggcttc ataccacctc agccagccgt gttataactt accgtttacc
     4141 aactacattt tttgtaacga accaaaaaac cctcaaaaga caagaccctc gaggtcgacg
     4201 aattcttaat taacatggca agctccaagg gtgaagagct atttactggg gttgtaccca
     4261 ttttggtaga actggacgga gatgtaaacg gacataaatt ctctgttaga ggtgagggcg
     4321 aaggcgatgc caccaatggt aaattgactc tgaagtttat atgcactacg ggtaaattac
     4381 ctgttccttg gccaacccta gtaacaactt tgacatatgg tgttcaatgt ttctcaagat
     4441 acccagacca tatgaaaagg catgatttct ttaaaagtgc tatgccagaa ggctacgtgc
     4501 aagagagaac tatctccttt aaggatgacg gtacgtataa aacacgagca gaagtgaaat
     4561 tcgaagggga tacactagtt aatcgcatcg aattaaaggg tatagacttt aaggaagatg
     4621 gtaatattct cggccataaa cttgagtata atttcaactc gcataatgtg tacattacag
     4681 ctgacaaaca aaagaacgga attaaagcga attttaaaat caggcacaac gtcgaagatg
     4741 ggtctgttca acttgccgat cattatcagc aaaacacccc tattggtgat ggtccagtct
     4801 tgttacccga taatcactac ttaagcacac agtctagatt gtcaaaagat ccgaatgaaa
     4861 agcgtgatca catggtttta ttggaatttg tcaccgctgc aggaataact cacggaatgg
     4921 acgagcttta taagcccggg gcggccgctt aaggcgcgcc tcatgtaatt agttatgtca
     4981 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     5041 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     5101 ttatatttca aatttttctt ttttttctgt acaaacgcgt gtacgcatgt aacattatac
     5161 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc agatctgagc
     5221 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     5281 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5341 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5401 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5461 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5521 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5581 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6301 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6361 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6421 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6481 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6541 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6601 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6661 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6721 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6781 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6841 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6901 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6961 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7021 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7081 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7141 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7201 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7261 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7321 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7381 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7441 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
