LOCUS       sequence-361476-        6305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bindingPCA plasmid - Contains a barcode landing pad and two 
            fragments of the murine methotrexate-resistant DHFR under the 
            expression of two CYC promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6305)
  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 6305)
  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: pGJJ001
FEATURES             Location/Qualifiers
     source          1..6305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(97..116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      910..1498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1399..1418
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1652..1669
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1786..1807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1822..1827,1828..1845,1846..1852)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     1913..1933
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1915..1933
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2577..2595)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      2607..2796
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      3672..4127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3759..3778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3969..3990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4258..5061)
                     /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(5062..5277)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5301..5320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5420..5442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5480..5498)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5539..6042
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6079..6183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6184..6252,6253..6305,1..739)
                     /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 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
       61 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      121 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      181 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      241 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      301 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      361 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      421 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      481 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      541 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      601 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      661 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      721 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      781 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      841 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      901 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      961 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1021 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1081 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1141 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1201 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1261 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1321 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1381 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1441 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1501 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1561 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1621 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1681 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1741 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     1801 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     1861 tgtgcgtacg actaagagcc taggcagcta tgaccatgat tacgccaagc gcgcaattaa
     1921 ccctcactaa agggaacaaa agctggagct catttggcga gcgttggttg gtggatcaag
     1981 cccacgcgta ggcaatcctc gagcagatcc gccaggcgtg tatatatagc gtggatggcc
     2041 aggcaacttt agtgctgaca catacaggca tatatatatg tgtgcgacga cacatgatca
     2101 tatggcatgc atgtgctctg tatgtatata aaactcttgt tttcttcttt tctctaaata
     2161 ttctttcctt atacattagg acctttgcag cataaattac tatacttcta tagacacgca
     2221 aacacaaata cacacactaa cggggctgct ctagaatgag taaagtagac atggtttgga
     2281 tagtcggagg cagttctgtt taccaggaag ccatgaatca accaggccac ctcagactct
     2341 ttgtgacaag gatcatgcag gaatttgaaa gtgacacgtt tttcccagaa attgacttgg
     2401 ggaaatataa acttctccca gaatacccag gcgtcctctc tgaggtccag gaggaaaaag
     2461 gcatcaagta taagtttgaa gtctacgaga agaaagacgg cggtggcggg tctggtggag
     2521 gagggtcagg tggtggcggc tcgggaggtg gagctagcta aaagcttatt agttatgtca
     2581 cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa
     2641 cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt
     2701 atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg
     2761 aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgact agtttaggat
     2821 ccgcctccac ctgacccacc tcctccagac ccgcctccgc ctgacccacc acctccggta
     2881 cccaattccg gttgttcaat aagtcttaag gcatcatcca aacttttggc aagaaaatga
     2941 gctcctcgtg gtggttcttt gagttctcta ctgagaacta tattaattct gtcctttaaa
     3001 ggtcgattct tctcaggaat ggagaaccag gttttcctac ccataatcac cagattctgt
     3061 ttaccttcca ctgaagaggt tgtggtcatt ctttggaagt acttggactc gttcctgagc
     3121 ggaggccagg gatagtctcc gttcttgcca atccccatat tttgggacac ggcgacgatg
     3181 cagttcaatg gtcgaaccat tctagagcag ccccgttagt gtgtgtattt gtgtttgcgt
     3241 gtctatagaa gtatagtaat ttatgctgca aaggtcctaa tgtataagga aagaatattt
     3301 agagaaaaga agaaaacaag agttttatat acatacagag cacatgcatg ccatatgatc
     3361 atgtgtcgtc gcacacatat atatatgcct gtatgtgtca gcactaaagt tgcctggcca
     3421 tccacgctat atatacacgc ctggcggatc tgctcgagga ttgcctacgc gtgggcttga
     3481 tccaccaacc aacgctcgcc aaatgagctc cagcttttgt tccctttagt gagcgtcgtg
     3541 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3601 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3661 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3721 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3781 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3841 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3901 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3961 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4021 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4081 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     4141 tttctcctta cgcatctgtg cggtatttca caccgcatag ggtaataact gatataatta
     4201 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     4261 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     4321 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     4381 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
     4441 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     4501 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
     4561 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     4621 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     4681 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
     4741 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
     4801 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     4861 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     4921 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     4981 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     5041 cttatatgta gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag
     5101 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     5161 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     5221 aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag
     5281 gtggtatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5341 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5401 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5461 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5521 ataatggttt cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat
     5581 gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt
     5641 ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag
     5701 gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca
     5761 gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt
     5821 gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag
     5881 agcaagataa aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa
     5941 acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt tatatattta
     6001 tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc
     6061 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6121 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6181 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6241 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6301 gcacg
//
