LOCUS       sequence-454432-        3691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SICLOPPS precursor for cyclo-CRVWCAR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3691)
  AUTHORS   Yang L, Zhang J, Andon JS, Li L, Wang T
  TITLE     Rapid discovery of cyclic peptide protein aggregation inhibitors by 
            continuous selection.
  JOURNAL   Nat Chem Biol. 2025 Jan 13. doi: 10.1038/s41589-024-01823-x.
  PUBMED    39806068
REFERENCE   2  (bases 1 to 3691)
  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: "Nat Chem 
            Biol. 2025 Jan 13. doi: 10.1038/s41589-024-01823-x."
COMMENT     Sequence Label: pLY090a
FEATURES             Location/Qualifiers
     source          1..3691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(80..625)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(160..179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(737..1819)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1781..1800
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1820..1897)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(2140..2145,2146..2161,2162..2168)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2176..2192
                     /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     2181..2203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             complement(2815..3630)
                     /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(2815..3630)
                     /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     3538..3557
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3622..3641
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 acggatcggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
       61 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
      121 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      181 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
      241 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
      301 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
      361 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
      421 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
      481 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
      541 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagatat
      601 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aaggagcccc atacgatata
      661 cgagaggaca tccgggcaaa tgcaacgaaa ggctcagtcg aaagactggg cctttcgttg
      721 cagacttagg gacccttcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      781 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
      841 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
      901 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
      961 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     1021 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     1081 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     1141 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     1201 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     1261 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     1321 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     1381 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     1441 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     1501 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     1561 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     1621 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     1681 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     1741 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     1801 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     1861 ccataccgcg aaaggttttg cgccattcga tggtgtcgcg caacgcaatt aatgtaagtt
     1921 agctcactca ttaggcacaa ttctcatgtt tgacagctta tcatcgactg cacggtgcac
     1981 caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat
     2041 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
     2101 acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa tcatcggctc
     2161 gtataatgtg tggaattgtg agcggataac aatttcacac aggaaacagc cagtccgttt
     2221 aggtgttttc acgagcactt caccaacaat aggtgcagta aggaggaaaa aaaaatggtt
     2281 aaagttatcg gtcgtcgttc cctcggagtg caaagaatat ttgatattgg tcttccccaa
     2341 gaccataatt ttctgctagc caatggggcg atcgcccaca actgtcgtgt gtggtgcgcc
     2401 cgctgtctca gtttcggcac cgaaatttta accgttgagt acggcccatt gcccattggc
     2461 aaaattgtga gtgaagaaat taattgttct gtgtacagtg ttgatccaaa agggagagtt
     2521 tacacccagg cgatcgccca atggcatgac cggggagagc aggaagtatt ggaatatgaa
     2581 ttggaagatg gttcagtaat ccgagctacc tctgaccacc gctttttaac caccgattat
     2641 caactgttgg cgatcgaaga aatttttgct aggcaactgg acttgttgac tttagaaaaa
     2701 aattaaatta acggcactgc gaaaaaaccc cgccgaagcg gggttttttg cgaaatataa
     2761 tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgtacg gatcttagaa
     2821 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2881 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     2941 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     3001 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     3061 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     3121 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     3181 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     3241 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     3301 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     3361 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     3421 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     3481 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     3541 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     3601 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     3661 agacagtttt attgttcatg aagagaaacc t
//
