LOCUS       sequence-117312-        9351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express an additional copy of Cdc24, integrated at TRP locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9351)
  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 9351)
  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-Cdc24
FEATURES             Location/Qualifiers
     source          1..9351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5898..5917)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5899..5917)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5924..5941)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5924..5940)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5933..5955)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6081..6536
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6168..6187)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6378..6399
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6892..6911)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7011..7033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7071..7089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7174..7278
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7279..7347,7348..8139)
                     /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(7497..7516)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8310..8898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8799..8818
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9052..9069
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9186..9207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9222..9227,9228..9245,9246..9252)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9260..9276
                     /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     9265..9287
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9284..9300
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9284..9300
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9321..9339
                     /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 cgatatggcg atccaaaccc gttttgcctc gggcacatct ttatccgatt tgaaaccaaa
     2281 accaagtgca acttccatct ccatacccat gcaaaatgtc atgaacaagc ctgtcacgga
     2341 acaggactca ctgttccata tatgcgcaaa catccggaaa agactggagg tgttacctca
     2401 actcaaacct tttttacaat tggcctacca atcgagcgag gttttgagtg aaaggcaatc
     2461 tcttttgcta tcccaaaagc agcatcagga actgctcaag tccaatggcg ctaaccggga
     2521 cagtagcgac ttggcaccaa ctttaaggtc tagctctatc tccacagcta ccagtctcat
     2581 gtcgatggaa ggtatatcat acacgaattc gaatccctcg gccaccccaa atatggagga
     2641 cactttactg acttttagta tgggtatttt gcccattacc atggattgcg accctgtgac
     2701 acaactatca cagctgtttc aacaaggtgc gcccctctgt atacttttca actctgtgaa
     2761 gccgcaattt aaattaccgg taatagcatc tgacgatttg aaagtctgta aaaaatccat
     2821 ttatgacttt atattgggct gcaagaaaca ctttgcattt aacgatgagg agcttttcac
     2881 tatatccgac gtttttgcca actctacttc ccagctggtc aaagtgctag aagtagtaga
     2941 aacgctaatg aattccagcc ctactatttt cccctctaag agtaagacac agcaaatcat
     3001 gaacgcagaa aaccaacacc gacatcagcc tcagcagtct tcgaagaagc ataacgagta
     3061 tgttaaaatt atcaaggaat tcgttgcaac ggaaagaaaa tatgttcacg atttggaaat
     3121 tttggataaa tatagacagc agttattaga cagcaatcta ataacgtctg aagagttgta
     3181 catgttgttc cctaatttgg gtgatgctat agattttcaa agaagatttc taatatcctt
     3241 ggaaataaat gctttagtag aaccttccaa gcaaagaatc ggggctcttt tcatgcattc
     3301 caaacatttt tttaagttgt atgagccttg gtctattggc caaaatgcag ccatcgaatt
     3361 tctctcttca actttgcaca agatgagggt tgatgaatcg cagcggttca taattaacaa
     3421 taaactggaa ttgcaatcct tcctttataa acccgtgcaa aggctttgta gatatcccct
     3481 gttggtcaaa gaattgcttg ctgaatcgag tgacgataat aatacgaaag aacttgaagc
     3541 tgctttagat atttctaaaa atattgcgag aagtatcaac gaaaatcaaa gaagaacaga
     3601 aaatcatcaa gtggtgaaga aactttatgg tagagtggtc aactggaagg gttatagaat
     3661 ttccaagttc ggtgagttat tatatttcga taaagtgttc atttcaacaa caaatagctc
     3721 ctcggaacct gaaagagaat ttgaggttta tctttttgaa aaaatcatca tccttttttc
     3781 agaggtagtg actaagaaat ctgcatcatc actaatcctt aagaagaaat cctcaacctc
     3841 agcatcaatc tccgcctcga acataacgga caacaatggc agccctcacc acagttacca
     3901 taagaggcat agcaatagta gtagcagtaa taatatccat ttatcttcgt cttcagcagc
     3961 ggcgataata cattccagta ccaatagtag tgacaacaat tccaacaatt catcatcatc
     4021 ctcattattc aagctgtccg ctaacgaacc taagctggat ctaagaggtc gaattatgat
     4081 aatgaatctg aatcaaatca taccgcaaaa caaccggtca ttaaatataa catgggaatc
     4141 cataaaagag caaggtaatt tccttttgaa attcaaaaat gaggaaacaa gagataattg
     4201 gtcatcgtgt ttacaacagt tgattcatga tctgaaaaat gagcagttta aggcaagaca
     4261 tcactcttca acatcgacga cttcatcgac agccaaatca tcttcaatga tgtcacccac
     4321 cacaactatg aatacaccga atcatcacaa cagccgccag acacacgata gtatggcttc
     4381 tttctcaagt tctcatatga aaagggtttc ggatgtcctg cctaaacgga ggaccacttc
     4441 atcaagtttc gaaagtgaaa ttaaatccat ttcagaaaat ttcaagaact ctattccaga
     4501 atcttccata ctcttcagga tatcatataa taacaactct aataatacct ctagtagcga
     4561 gatcttcaca cttttggtag aaaaagtttg gaattttgac gacttgataa tggcgatcaa
     4621 ttctaaaatt tcgaatacac ataataacaa catttcacca atcaccaaga tcaaatatca
     4681 ggacgaagat ggggattttg ttgtgttagg tagcgatgaa gattggaatg ttgctaaaga
     4741 aatgttggcg gaaaacaatg agaaattctt gaacattcgt ctgtattgag cggccgccac
     4801 cgcggtggag ctctaagcaa atagctaaat tatatacgaa ttaatattat gattaagtgt
     4861 ttacgtgagt gcgatatttt tattactatc ttatacagtt gtatatactc tataaaatga
     4921 gttgtctatt aattaacgcg atgaatgctt tctgggttta cctctccaac aactctagtt
     4981 tacttctcaa tacattcaat tgtatttgat ttgtcaatac ttcatcatta atcaattcta
     5041 tagttttgtt tttctcgttt atttccaaat ttaatgcatc aattttatta ttcaatttgt
     5101 cgttgatttt ggttaatgat tttatggttt gatctctggc attgattgtt tgtgttagtt
     5161 tttcattatt gataattaaa ttatttaagt tagttatcaa ctcggtgttt tcaagtttca
     5221 agttttcaat ttctttagag tttattagat ttgtcaaagt ttctgaattg cttgattggt
     5281 cctgtagaag agtatttgtt gttgtggata attgattcaa tttttgagac aattgctgga
     5341 aggcgttgaa atatctagca tcaatctcat ggtttttttc ccgagagtct cgtagattca
     5401 attgttttaa tatatcttgg gaccactctt gatttgaact catggaaatt aaactgggtg
     5461 ttgtgttgtg gtgtaatgat tgtaccccct ttgcttataa ttgtgtggtt ccgatgctga
     5521 cttactgggt attatatgtg tgcccaatag aaagagaaca attgacccgg ttattgcaag
     5581 gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga aatacttggt
     5641 tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg attacggcat
     5701 tgatatcgtc caactgcatg gagatgagtc gtggcgagaa taccaagagt tcctcggttt
     5761 gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca gtgcagcttc
     5821 acagaaacct cattcgttta ttcccttgtt tgattcagaa ggccggccat ttaaatgttt
     5881 aaacggccac gctggccccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     5941 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6001 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6061 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6121 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6181 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6241 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6301 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6361 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6421 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6481 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     6541 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg
     6601 atataattaa attgaagctc taatttgtga gtttagtatt aggatctcta cccttggcga
     6661 aaagtcctct gccaacaatg atgatatctg atccaccact tacaacttcg tcgacggttc
     6721 tgtactgctg acccaatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     6781 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
     6841 aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg tgcactctca gtacaatctg
     6901 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     6961 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7021 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     7081 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt cggcgcgccg
     7141 aggacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7201 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7261 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7321 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7381 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7441 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7501 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7561 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7621 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7681 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7741 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7801 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7861 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7921 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7981 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8041 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8101 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8161 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8221 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8281 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8341 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8401 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8461 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8521 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8581 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     8641 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8701 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8761 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8821 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8881 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8941 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     9001 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     9061 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     9121 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     9181 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     9241 gctcctatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     9301 catgattacg ccaagctcgg aattaaccct cactaaaggg aacaaaagct g
//
