LOCUS       sequence-275756-        7324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in fission yeast from promoter pact1, integrated at lys3 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7324)
  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 7324)
  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: pAV0662
FEATURES             Location/Qualifiers
     source          1..7324
                     /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"
     terminator      4790..5039
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4806..4824)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5064..5084)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5064..5082)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5103..5119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5103..5119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5116..5138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5127..5143
                     /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(5151..5157,5158..5175,5176..5181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5196..5217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5334..5351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5505..6093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5585..5604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6264..7055,7056..7124))
                     /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     6887..6906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7125..7229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7297..7315
                     /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 gggcccattt aaatgatcta cgataatgag acggtgtttg ggttgttttt actgaatttt
     3241 tcttttattt tatgacatct taatttttat gttaatgatt attcttatac ttttatctat
     3301 ctttgattaa aactaatatt cattcttttg atcactaatt atttttatag tatacgattc
     3361 aacccaacta taatacattg tatcaagaga aatttttaat tagatgtaat ttaatagcca
     3421 tttgataaaa aaaaattacc tttttttttg ctagtgttaa attattgatc ataacaatgt
     3481 cgaatgaata ctaataaaat agggacacgc gagttgcaaa tacttgaaat acctttgaag
     3541 atttgtctgt gaattttgat aagattttta tgaccatctc ttttatattc gatatcgtga
     3601 actttcattt tcttttgttt tttgcttttc aagttaaata acgcggaaat tatatttcct
     3661 tatatgttta taagagcttt taaaggattt gctatatcgc ttacagtact acaaccggtt
     3721 gctcaatgtt atccgtttcc gcggtcttct tccgtgcgca ttctgccgtg aagtgctaac
     3781 gctgtgtgtg gctttatcaa aaaagtggct caaatatagc aatttcatac atatttaaac
     3841 acccattata cttaaatgta tattttaaaa tatataagag attttcactg tataactgtg
     3901 tatcttatct atatgtaatc aacggcttca taccacctca gccagccgtg ttataactta
     3961 ccgtttacca actacatttt ttgtaacgaa ccaaaaaacc ctcaaaagac aagaccctcg
     4021 aggtcgacga attcttaatt aacatggcaa gctccaaggg tgaagagcta tttactgggg
     4081 ttgtacccat tttggtagaa ctggacggag atgtaaacgg acataaattc tctgttagag
     4141 gtgagggcga aggcgatgcc accaatggta aattgactct gaagtttata tgcactacgg
     4201 gtaaattacc tgttccttgg ccaaccctag taacaacttt gacatatggt gttcaatgtt
     4261 tctcaagata cccagaccat atgaaaaggc atgatttctt taaaagtgct atgccagaag
     4321 gctacgtgca agagagaact atctccttta aggatgacgg tacgtataaa acacgagcag
     4381 aagtgaaatt cgaaggggat acactagtta atcgcatcga attaaagggt atagacttta
     4441 aggaagatgg taatattctc ggccataaac ttgagtataa tttcaactcg cataatgtgt
     4501 acattacagc tgacaaacaa aagaacggaa ttaaagcgaa ttttaaaatc aggcacaacg
     4561 tcgaagatgg gtctgttcaa cttgccgatc attatcagca aaacacccct attggtgatg
     4621 gtccagtctt gttacccgat aatcactact taagcacaca gtctagattg tcaaaagatc
     4681 cgaatgaaaa gcgtgatcac atggttttat tggaatttgt caccgctgca ggaataactc
     4741 acggaatgga cgagctttat aagcccgggg cggccgctta aggcgcgcct catgtaatta
     4801 gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga
     4861 gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa
     4921 gaacgttatt tatatttcaa atttttcttt tttttctgta caaacgcgtg tacgcatgta
     4981 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca
     5041 gatctgagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     5101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5161 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5281 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5341 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5401 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5701 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5941 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7261 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7321 cgtc
//
