LOCUS       sequence-126394-        8442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express an additional copy of Bem1, integrated at TRP locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8442)
  AUTHORS   Jost AP, Weiner OD
  TITLE     Probing Yeast Polarity with Acute, Reversible, Optogenetic 
            Inhibition of Protein Function.
  JOURNAL   ACS Synth Biol. 2015 Jun 2.
  PUBMED    26035630
REFERENCE   2  (bases 1 to 8442)
  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: "ACS Synth 
            Biol. 2015 Jun 2."
COMMENT     Sequence Label: pCyc-Bem1
FEATURES             Location/Qualifiers
     source          1..8442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4989..5008)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4990..5008)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5015..5032)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5015..5031)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5024..5046)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5172..5627
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5259..5278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5469..5490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5983..6002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6102..6124
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6162..6180)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6265..6369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6370..6438,6439..7230)
                     /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     complement(6588..6607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7401..7989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7890..7909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8143..8160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8277..8298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8313..8318,8319..8336,8337..8343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8351..8367
                     /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)."
     primer_bind     8356..8378
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8375..8391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8375..8391
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8412..8430
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccata tttctaccaa tctctcaaca
       61 ctgagtaatg gtagttataa gaaagagacc gagttaggga cagttagagg cggtggagat
      121 attccttatg gcatgtctgg cgatgataaa acttttcaaa cggcagcccc gatctaaaag
      181 agctgacagg gaaatggtca gaaaaagaaa cgtgcacccg cccgtctgga cgcgccgctc
      241 acccgcacgg cagagaccaa tcagtaaaaa tcaacggtta acgacattac tatatatata
      301 atataggaag catttaatag aacagcatcg taatatatgt gtactttgca gttatgacgc
      361 cagatggcag tagtggaaga tattctttat tgaaaaatag cttgtcacct tacgtacaat
      421 cttgatccgg agcccgtcgc tttgagagaa atcagaagat tccaaaagtc tactgaactt
      481 ttgatcagaa agttgccttt ccaaagacta gtcagagaaa tcgcccaaga tttcaagacc
      541 gatctaagat tccaatcctc tgccatcggt gctctacagg aatccgttga agcttacttg
      601 gtctctttgt tcgaagacac caacttggcc gctatccacg ctaagcgtgt taccatccaa
      661 aagaaggata tcaagttggc tagaagattg agaggtgaaa gatcctaagc acacagggat
      721 tgttgtacaa ttacttttta tgattttaga tacacacaaa tcttttctac gattaccttt
      781 catttatttt cgtttcatgt tttattaatc attctttttg tatatttcac atcgggtggg
      841 gtttttatat taaattttaa taacataatc ttttaaagtt tctactttct atcctgtata
      901 ttatttccat atctatatct atattttcaa gcttgttcac atccttatcg gaattgttcc
      961 agcgtttttg tacagaaaat gatgttatgg taacgagcta tatcagacct actagtctta
     1021 tagagctgta cagagtcttg gaaacataac caacacacca acaagatgtc atttgattcg
     1081 ttactcgaca agaatgataa gctggtaaaa gtttgcggga ttcaaaccgt cgaggctgcc
     1141 gaaactgcgc ttcaagcagg cgctgatttg atagggatca tatgtgtccc caacaggaag
     1201 cggactatcg agagcgctgt ggctcgtgaa atatccaaat tgattcacaa atcaggtact
     1261 acaaagctgg tgggggtgtt caggaatcaa tctgttgagg acgtacatcg gctttctgag
     1321 gaatatgacc ttgacataat ccaattacat ggtgatgaat catggccaga gtactataac
     1381 gtcattaaga aaccaataat caaaagagtc atattcccta gagatgtcga tgttgtaaca
     1441 caagtgtgtc aaagaaaacc cttggtatgt ctaccattgt tcgactccga ggcaggcggt
     1501 acaggtgaaa agctggactg gtccagtatt tcctcctggg caagcgaaca gaataacgta
     1561 ttttatatac tagcgggtgg actaactgca gaaaacgtca tggaggcggt ggggttgcct
     1621 ggtgtaattg gtgttgacgt ctcaggtggt gtagagactg acggcgtcaa agacaatgac
     1681 aagataatta aatatgtaca aaatgcaaag aaacaatgaa ttacgaagag aagtctcaat
     1741 gaggttttac aaacaatcct ataatacata tatgccactt agatatactt tgaatactga
     1801 atatactgac tcattatatt ctttaattgt agtttctcaa aaatattcat acaggttagc
     1861 tatgaattca tgctataatt tccattttgt tcatgtaaga tacacatagg gataaaatgt
     1921 gataactaat cagcggtacc gggcccgagc agatccgcca ggcgtgtata tatagcgtgg
     1981 atggccaggc aactttagtg ctgacacata caggcatata tatatgtgtg cgacgacaca
     2041 tgatcatatg gcatgcatgt gctctgtatg tatataaaac tcttgttttc ttcttttctc
     2101 taaatattct ttccttatac attaggacct ttgcagcata aattactata cttctataga
     2161 cacacaaaca caaatacaca cactaaatta ataaagcaca aggcaggtgg tcgacggtat
     2221 cgatatgctg aaaaacttca aactctcaaa aagagatagt aatgggtcga agggcagaat
     2281 tacatccgca gatatttcca caccttctca tgataatggg agtgttataa agcatattaa
     2341 gacagtacca gtgaggtacc tttcttcatc ttccacccca gtgaaaagcc aacgggactc
     2401 ttcaccaaaa aacagacata attctaaaga tattacttct ccagagaaag ttataaaagc
     2461 caaatacagt tatcaggcac aaacttcaaa ggaactatct ttcatggaag gtgaatttat
     2521 ttatgtatct ggagatgaga aggattggta taaagcttca aatccttcta ctggaaaaga
     2581 gggtgttgtc cccaaaacct attttgaagt gtttgataga actaaacctt cctctgtaaa
     2641 tggatccaat agttcaagtc ggaaggttac aaatgattca ttaaacatgg gatcgttata
     2701 cgccattgtt ttatatgact ttaaggcaga aaaagctgat gagttgacaa cttatgtggg
     2761 agagaatttg tttatttgcg cccaccataa ctgtgaatgg ttcattgcta agccaattgg
     2821 tcgacttgga gggcccggcc ttgttcctgt tgggtttgtt agcatcatag atattgccac
     2881 ggggtacgca acaggtaatg acgtaataga agacatcaag tcagttaacc taccaactgt
     2941 tcaagaatgg aaaagcaaca ttgcaagata caaagctagt aatattagcc ttggttctgt
     3001 ggagcagcag cagcagcaat ccattacaaa acctcagaac aaaagcgcaa agctggttga
     3061 cggtgaatta ttagtgaaag catccgttga atcctttggg ttagaagatg agaagtattg
     3121 gtttttggtt tgctgtgaat tatctaacgg taagacgagg caactgaaaa gatactatca
     3181 agatttctac gatctgcagg ttcaactttt ggatgcgttc ccagcagagg ccggtaagtt
     3241 gagggatgca ggtggacaat ggtccaaacg tataatgccc tatatccctg gacccgttcc
     3301 atacgttacc aatagtataa caaaaaaaag aaaggaggac ttgaatatat atgtggcaga
     3361 cctagtgaat cttcccgatt atatatctcg ctcagaaatg gtgcattcct tattcgttgt
     3421 tttgaataat ggtttcgaca gagaatttga aagagacgaa aatcaaaaca atattaaaac
     3481 attacaagaa aacgatacag caacctttgc aacagcatct caaacttcta attttgcctc
     3541 aactaaccaa gacaacactt taactggcga agacttgaag ttaaacaaga aactctcaga
     3601 tttatcttta tcgggctcca agcaggctcc agcccaatca acttctggat tgaaaacaac
     3661 taaaattaaa ttttattaca aagacgatat ttttgcttta atgctgaagg gtgacacaac
     3721 ctataaagaa cttcgtagta aaatcgctcc aaggattgat acagataatt ttaaattaca
     3781 aaccaaatta tttgatggta gcggggagga aattaagact gattcacaag tcagtaacat
     3841 aatccaagcc aaactgaaaa tttccgttca cgatatttga gcggccgcca ccgcggtgga
     3901 gctctaagca aatagctaaa ttatatacga attaatatta tgattaagtg tttacgtgag
     3961 tgcgatattt ttattactat cttatacagt tgtatatact ctataaaatg agttgtctat
     4021 taattaacgc gatgaatgct ttctgggttt acctctccaa caactctagt ttacttctca
     4081 atacattcaa ttgtatttga tttgtcaata cttcatcatt aatcaattct atagttttgt
     4141 ttttctcgtt tatttccaaa tttaatgcat caattttatt attcaatttg tcgttgattt
     4201 tggttaatga ttttatggtt tgatctctgg cattgattgt ttgtgttagt ttttcattat
     4261 tgataattaa attatttaag ttagttatca actcggtgtt ttcaagtttc aagttttcaa
     4321 tttctttaga gtttattaga tttgtcaaag tttctgaatt gcttgattgg tcctgtagaa
     4381 gagtatttgt tgttgtggat aattgattca atttttgaga caattgctgg aaggcgttga
     4441 aatatctagc atcaatctca tggttttttt cccgagagtc tcgtagattc aattgtttta
     4501 atatatcttg ggaccactct tgatttgaac tcatggaaat taaactgggt gttgtgttgt
     4561 ggtgtaatga ttgtaccccc tttgcttata attgtgtggt tccgatgctg acttactggg
     4621 tattatatgt gtgcccaata gaaagagaac aattgacccg gttattgcaa ggaaaatttc
     4681 aagtcttgta aaagcatata aaaatagttc aggcactccg aaatacttgg ttggcgtgtt
     4741 tcgtaatcaa cctaaggagg atgttttggc tctggtcaat gattacggca ttgatatcgt
     4801 ccaactgcat ggagatgagt cgtggcgaga ataccaagag ttcctcggtt tgccagttat
     4861 taaaagactc gtatttccaa aagactgcaa catactactc agtgcagctt cacagaaacc
     4921 tcattcgttt attcccttgt ttgattcaga aggccggcca tttaaatgtt taaacggcca
     4981 cgctggcccc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
     5041 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     5101 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     5161 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5221 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5281 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5341 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5401 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5461 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5521 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5581 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     5641 tttctcctta cgcatctgtg cggtatttca caccgcatag ggtaataact gatataatta
     5701 aattgaagct ctaatttgtg agtttagtat taggatctct acccttggcg aaaagtcctc
     5761 tgccaacaat gatgatatct gatccaccac ttacaacttc gtcgacggtt ctgtactgct
     5821 gacccaatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     5881 gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa
     5941 aaaaaaatga tgaattgaaa aggtggtatg gtgcactctc agtacaatct gctctgatgc
     6001 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6061 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6121 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     6181 tttataggtt aatgtcatga taataatggt ttcttagacg tcggcgcgcc gaggacgtca
     6241 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6301 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6361 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6421 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6481 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6541 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6601 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6661 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6721 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6781 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6841 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6901 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6961 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7021 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7081 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7141 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7201 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7261 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7321 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7381 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7441 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7501 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7561 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7621 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7681 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7741 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7801 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7861 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7921 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7981 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8041 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8101 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8161 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8221 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8281 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcctatg
     8341 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8401 gccaagctcg gaattaaccc tcactaaagg gaacaaaagc tg
//
