LOCUS       sequence-270894-        4000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PSE1 experimental evolution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4000)
  AUTHORS   Stiffler MA, Poelwijk FJ, Brock KP, Stein RR, Riesselman A, Teyra J,
            Sidhu SS, Marks DS, Gauthier NP, Sander C
  TITLE     Protein Structure from Experimental Evolution.
  JOURNAL   Cell Syst. 2020 Jan 22;10(1):15-24.e5. doi: 
            10.1016/j.cels.2019.11.008. Epub 2019 Dec 11.
  PUBMED    31838147
REFERENCE   2  (bases 1 to 4000)
  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: 
            "10.1016/j.cels.2019.11.008"; journalName: "Cell Syst"; date: 
            "2020-01-22- 22"; volume: "10"; issue: "1"; pages: "15-24.e5"
COMMENT     Sequence Label: pBR322_KanR_AgeI_AvrII_PSE1
FEATURES             Location/Qualifiers
     source          1..4000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..36)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        104..208
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1264..1852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1753..1772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2006..2023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2038..2178
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2104..2123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2264..2286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2280..2471)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3100..3915)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3100..3915)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3823..3842
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(join(3963..3968,3969..3985,3986..3991))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gcctaccggt ttttgctagt agttcaaagt ttcagcaagt
      301 tgaacaagac gttaaggcaa ttgaagtttc tctttctgct cgtataggtg tttccgttct
      361 tgatactcaa aatggagaat attgggatta caatggcaat cagcgcttcc cgttaacaag
      421 tacttttaaa acaatagctt gcgctaaatt actatatgat gctgagcaag gaaaagttaa
      481 tcccaatagt acagtcgaga ttaagaaagc agatcttgtg acctattccc ctgtaataga
      541 aaagcaagta gggcaggcaa tcacactcga tgatgcgtgc ttcgcaacta tgactacaag
      601 tgataatact gcggcaaata tcatcctaag tgctgtaggt ggccccaaag gcgttactga
      661 ttttttaaga caaattgggg acaaagagac tcgtctagac cgtattgagc ctgatttaaa
      721 tgaaggtaag ctcggtgatt tgagggatac gacaactcct aaggcaatag ccagtacttt
      781 gaataaattt ttatttggtt ccgcgctatc tgaaatgaac cagaaaaaat tagagtcttg
      841 gatggtgaac aatcaagtca ctggtaattt actacgttca gtattgccgg cgggatggaa
      901 cattgcggat cgctcaggtg ctggcggatt tggtgctcgg agtattacag cagttgtgtg
      961 gagtgagcat caagccccaa ttattgtgag catctatcta gctcaaacac aggcttcaat
     1021 ggcagagcga aatgatgcga ttgttaaaat tggtcattca atttttgacg tttatacatc
     1081 acagtcgcgc taactgtcag acctaggtta ctcatatata ctttagattg atttaaaact
     1141 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1381 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     1921 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     1981 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2041 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     2101 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     2161 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2221 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2281 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     2341 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     2401 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     2461 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     2521 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     2581 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     2641 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     2701 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     2761 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     2821 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     2881 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     2941 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     3001 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     3061 ccaattcttg gagtggtgta tccgctcatg aattaattct tagaaaaact catcgagcat
     3121 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     3181 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     3241 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     3301 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     3361 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     3421 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     3481 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     3541 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     3601 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     3661 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     3721 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     3781 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     3841 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg
     3901 ttgaatatgg ctcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc
     3961 gcattaaagc ttatcgatga taagctgtca aacatgagaa
//
