LOCUS       sequence-402862-        7538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human c-Src kinase from a GAL10 promoter with a URA3 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7538)
  AUTHORS   Hong SJ, Resnick SJ, Iketani S, Cha JW, Albert BA, Fazekas CT, Chang
            CW, Liu H, Dagan S, Abagyan MR, Fajtova P, Culbertson B, Brace B, 
            Reddem ER, Forouhar F, Glickman JF, Balkovec JM, Stockwell BR, 
            Shapiro L, O'Donoghue AJ, Sabo Y, Freundlich JS, Ho DD, Chavez A
  TITLE     A multiplex method for rapidly identifying viral protease 
            inhibitors.
  JOURNAL   Mol Syst Biol. 2025 Feb;21(2):158-172. doi: 
            10.1038/s44320-024-00082-1. Epub 2025 Jan 6.
  PUBMED    39762652
REFERENCE   2  (bases 1 to 7538)
  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"; doi: 
            "10.1038/s44320-024-00082-1"; journalName: "Mol Syst Biol"; date: 
            "2025-02"; volume: "21"; issue: "2"; pages: "158-172"
COMMENT     Sequence Label: pAG416GAL10-c-Src
FEATURES             Location/Qualifiers
     source          1..7538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1108..1549)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(1114..1137)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1432..1549
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     protein_bind    1827..1851
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3526..3542)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3527..3543)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3545..3792
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3561..3579)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3810..3829)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3811..3829)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3839..3856)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3839..3855)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3848..3870)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3996..4451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4083..4102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4293..4314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4582..5385)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5386..5601)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5625..5644)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5744..5766
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5804..5822)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5863..6366
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6403..6507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6508..6576,6577..7368)
                     /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(6726..6745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcta tgaccatgat tacgccaagc ggggtgtgtt
     1081 gttggtgggt tgggtgtata gttttttctc cttgacgtta aagtatagag gtatattaac
     1141 aattttttgt tgatactttt atgacatttg aataagaagt aatacaaact gaaaatgttg
     1201 aaagtattag ttaaagtggt tatgcagctt ttccatttat atatctgtta atagatcaaa
     1261 aatcatcgct tcgctgatta attaccccag aaataaggct aaaaaactaa tcgcattatc
     1321 atcctatggt tgttaatttg attcgttaat ttgaaggttt gtggggccag gttactgcca
     1381 atttttcctc ttcataacca taaaagctag tattgtagaa tctttattgt tcggagcagt
     1441 gcggcgcgag gcacatctgc gtttcaggaa cgcgaccggt gaagacgagg acgcacggag
     1501 gagagtcttc cgtcggaggg ctgtcgcccg ctcggcggct tctaatccgt acttcaatat
     1561 agcaatgagc agttaagcgt attactgaaa gttccaaaga gaaggttttt ttaggctaag
     1621 ataatggggc tctttacatt tccacaacat ataagtaaga ttagatatgg atatgtatat
     1681 ggtggtaatg ccatgtaata tgattattaa acttctttgc gtccatccaa aaaaaaagta
     1741 agaatttttg aaaattcaat ataaatgtct aaaggtgaag aattattcag aattccagct
     1801 gaccaccgac tagtggatcc cccatcacaa gtttgtacaa aaaagcaggc tacaaaatgg
     1861 gtagcaacaa gagcaagccc aaggatgcca gccagcggcg ccgcagcctg gagcccgccg
     1921 agaacgtgca cggcgctggc gggggcgctt tccccgcctc gcagaccccc agcaagccag
     1981 cctcggccga cggccaccgc ggccccagcg cggccttcgc ccccgcggcc gccgagccca
     2041 agctgttcgg aggcttcaac tcctcggaca ccgtcacctc cccgcagagg gcgggcccgc
     2101 tggccggtgg agtgaccacc tttgtggccc tctatgacta tgagtctagg acggagacag
     2161 acctgtcctt caagaaaggc gagcggctcc agattgtcaa caacacagag ggagactggt
     2221 ggctggccca ctcgctcagc acaggacaga caggctacat ccccagcaac tacgtggcgc
     2281 cctccgactc catccaggct gaggagtggt attttggcaa gatcaccaga cgggagtcag
     2341 agcggttact gctcaatgca gagaacccga gagggacctt cctcgtgcga gaaagtgaga
     2401 ccacgaaagg tgcctactgc ctctcagtgt ctgacttcga caacgccaag ggcctcaacg
     2461 tgaagcacta caagatccgc aagctggaca gcggcggctt ctacatcacc tcccgcaccc
     2521 agttcaacag cctgcagcag ctggtggcct actactccaa acacgccgat ggcctgtgcc
     2581 accgcctcac caccgtgtgc cccacgtcca agccgcagac tcagggcctg gccaaggatg
     2641 cctgggagat ccctcgggag tcgctgcggc tggaggtcaa gctgggccag ggctgctttg
     2701 gcgaggtgtg gatggggacc tggaacggta ccaccagggt ggccatcaaa accctgaagc
     2761 ctggcacgat gtctccagag gccttcctgc aggaggccca ggtcatgaag aagctgaggc
     2821 atgagaagct ggtgcagttg tatgctgtgg tttcagagga gcccatttac atcgtcacgg
     2881 agtacatgag caaggggagt ttgctggact ttctcaaggg ggagacaggc aagtacctgc
     2941 ggctgcctca gctggtggac atggctgctc agatcgcctc aggcatggcg tacgtggagc
     3001 ggatgaacta cgtccaccgg gaccttcgtg cagccaacat cctggtggga gagaacctgg
     3061 tgtgcaaagt ggccgacttt gggctggctc ggctcattga agacaatgag tacacggcgc
     3121 ggcaaggtgc caaattcccc atcaagtgga cggctccaga agctgccctc tatggccgct
     3181 tcaccatcaa gtcggacgtg tggtccttcg ggatcctgct gactgagctc accacaaagg
     3241 gacgggtgcc ctaccctggg atggtgaacc gcgaggtgct ggaccaggtg gagcggggct
     3301 accggatgcc ctgcccgccg gagtgtcccg agtccctgca cgacctcatg tgccagtgct
     3361 ggcggaagga gcctgaggag cggcccacct tcgagtacct gcaggccttc ctggaggact
     3421 acttcacgtc caccgagccc cagtaccagc ccggggagaa cctctaaaac ccagctttct
     3481 tgtactaagt ggtgatgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct
     3541 cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
     3601 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     3661 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     3721 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     3781 gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     3841 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3901 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3961 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     4021 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4081 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4141 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4201 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4261 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4321 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4381 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4441 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4501 atagggtaat aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc
     4561 atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca
     4621 gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc
     4681 tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg
     4741 ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc
     4801 gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt
     4861 gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt
     4921 gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc
     4981 ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc
     5041 ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc
     5101 agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg
     5161 cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca
     5221 aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa
     5281 tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg
     5341 actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt
     5401 cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc
     5461 aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt
     5521 ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata
     5581 aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat ctgctctgat
     5641 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5701 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5761 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     5821 tttttatagg ttaatgtcat gataataatg gtttcttaga cggatcgctt gcctgtaact
     5881 tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt
     5941 atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga
     6001 atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata
     6061 tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa
     6121 aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg
     6181 gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc
     6241 tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact
     6301 ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca
     6361 cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6421 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6481 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6541 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6601 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6661 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6721 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6781 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6841 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6901 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6961 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     7021 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     7081 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7141 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7201 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7261 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7321 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7381 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7501 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
