LOCUS       sequence-361478-        6671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  abundancePCA plasmid - Contains a barcode landing pad and two 
            fragments of the murine methotrexate-resistant DHFR, the expression 
            of one driven by the CYC promoter and the other by the GPD 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  AUTHORS   Faure AJ, Domingo J, Schmiedel JM, Hidalgo-Carcedo C, Diss G, Lehner
            B
  TITLE     Mapping the energetic and allosteric landscapes of protein binding 
            domains.
  JOURNAL   Nature. 2022 Apr;604(7904):175-183. doi: 10.1038/s41586-022-04586-4.
            Epub 2022 Apr 6.
  PUBMED    35388192
REFERENCE   2  (bases 1 to 6671)
  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/s41586-022-04586-4"; journalName: "Nature"; date: 
            "2022-04"; volume: "604"; issue: "7904"; pages: "175-183"
COMMENT     Sequence Label: pGJJ045
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      919..1507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1408..1427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1661..1678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1795..1816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1831..1836,1837..1854,1855..1861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     1922..1942
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1924..1942
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2586..2604)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      2616..2805
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(3230..3873)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(3313..3336)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     rep_origin      4047..4502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4134..4153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4344..4365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4633..5436)
                     /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(5437..5652)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5676..5695)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5795..5817
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5855..5873)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5914..6417
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6454..6558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6559..6627,6628..6671,1..748)
                     /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"
ORIGIN
        1 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
       61 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      121 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      421 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      601 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1621 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1681 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1741 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     1801 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     1861 gtgtggaatt gtgcgtacga ctaagagcct aggcagctat gaccatgatt acgccaagcg
     1921 cgcaattaac cctcactaaa gggaacaaaa gctggagctc atttggcgag cgttggttgg
     1981 tggatcaagc ccacgcgtag gcaatcctcg agcagatccg ccaggcgtgt atatatagcg
     2041 tggatggcca ggcaacttta gtgctgacac atacaggcat atatatatgt gtgcgacgac
     2101 acatgatcat atggcatgca tgtgctctgt atgtatataa aactcttgtt ttcttctttt
     2161 ctctaaatat tctttcctta tacattagga cctttgcagc ataaattact atacttctat
     2221 agacacgcaa acacaaatac acacactaac ggggctgctc tagaatgagt aaagtagaca
     2281 tggtttggat agtcggaggc agttctgttt accaggaagc catgaatcaa ccaggccacc
     2341 tcagactctt tgtgacaagg atcatgcagg aatttgaaag tgacacgttt ttcccagaaa
     2401 ttgacttggg gaaatataaa cttctcccag aatacccagg cgtcctctct gaggtccagg
     2461 aggaaaaagg catcaagtat aagtttgaag tctacgagaa gaaagacggc ggtggcgggt
     2521 ctggtggagg agggtcaggt ggtggcggct cgggaggtgg agctagctaa aagcttatta
     2581 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     2641 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     2701 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     2761 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgacta
     2821 gtttaggatc cgcctccacc tgacccacct cctccagacc cgcctccgcc tgacccacca
     2881 cctccggtac ccaattccgg ttgttcaata agtcttaagg catcatccaa acttttggca
     2941 agaaaatgag ctcctcgtgg tggttctttg agttctctac tgagaactat attaattctg
     3001 tcctttaaag gtcgattctt ctcaggaatg gagaaccagg ttttcctacc cataatcacc
     3061 agattctgtt taccttccac tgaagaggtt gtggtcattc tttggaagta cttggactcg
     3121 ttcctgagcg gaggccaggg atagtctccg ttcttgccaa tccccatatt ttgggacacg
     3181 gcgacgatgc agttcaatgg tcgaaccatt ctagagcagc cccgatccgt cgaaactaag
     3241 ttctggtgtt ttaaaactaa aaaaaagact aactataaaa gtagaattta agaagtttaa
     3301 gaaatagatt tacagaatta caatcaatac ctaccgtctt tatatactta ttagtcaagt
     3361 aggggaataa tttcagggaa ctggtttcaa cctttttttt cagctttttc caaatcagag
     3421 agagcagaag gtaatagaag gtgtaagaaa atgagataga tacatgcgtg ggtcaattgc
     3481 cttgtgtcat catttactcc aggcaggttg catcactcca ttgaggttgt gcccgttttt
     3541 tgcctgtttg tgcccctgtt ctctgtagtt gcgctaagag aatggaccta tgaactgatg
     3601 gttggtgaag aaaacaatat tttggtgctg ggattctttt tttttctgga tgccagctta
     3661 aaaagcgggc tccattatat ttagtggatg ccaggaataa actgttcacc cagacaccta
     3721 cgatgttata tattctgtgt aacccgcccc ctattttggg catgtacggg ttacagcaga
     3781 attaaaaggc taattttttg actaaataaa gttaggaaaa tcactactat taattattta
     3841 cgtattcttt gaaatggcga gtattgataa tgataaactg agctccagct tttgttccct
     3901 ttagtgagcg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3961 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4021 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4081 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4141 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4201 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4261 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4321 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4381 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4441 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     4501 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa
     4561 taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta
     4621 taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt
     4681 tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac
     4741 aataataatg tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa
     4801 tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc
     4861 atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt
     4921 cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa
     4981 ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa
     5041 accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc
     5101 tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt
     5161 tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt
     5221 gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg
     5281 acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca
     5341 caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg
     5401 agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt
     5461 ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca
     5521 tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga
     5581 gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa
     5641 tgatgaattg aaaaggtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5701 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5761 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5821 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5881 gttaatgtca tgataataat ggtttcttag acggatcgct tgcctgtaac ttacacgcgc
     5941 ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg
     6001 ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa
     6061 aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat
     6121 tagacaagaa aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa
     6181 tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata
     6241 cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt
     6301 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
     6361 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
     6421 aaggacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6481 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6541 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6601 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6661 gaagatcagt t
//
