LOCUS       sequence-454431-        3691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SICLOPPS precursor for cyclo-CRCVFSG.
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: pLY071f
FEATURES             Location/Qualifiers
     source          1..3691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(72..617)
                     /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(152..171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(729..1811)
                     /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     1773..1792
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1812..1889)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(2132..2137,2138..2153,2154..2160)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2168..2184
                     /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     2173..2195
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             complement(2807..3622)
                     /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(2807..3622)
                     /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     3530..3549
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3614..3633
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
       61 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
      121 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
      181 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
      241 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
      301 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
      361 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
      421 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
      481 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
      541 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagat attacgcgca
      601 gaccaaaacg atctcaagaa gatcatctta ttaaggagcc ccatacgata tacgagagga
      661 catccgggca aatgcaacga aaggctcagt cgaaagactg ggcctttcgt tgcagactta
      721 gggacccttc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
      841 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
      901 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
      961 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     1021 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     1081 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     1141 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     1201 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     1261 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     1321 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     1381 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     1441 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     1501 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     1561 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     1621 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     1681 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     1741 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     1801 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     1861 cgaaaggttt tgcgccattc gatggtgtcg cgcaacgcaa ttaatgtaag ttagctcact
     1921 cattaggcac aattctcatg tttgacagct tatcatcgac tgcacggtgc accaatgctt
     1981 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
     2041 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
     2101 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
     2161 tgtggaattg tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt
     2221 tcacgagcac ttcaccaaca ataggtgcag taaggaggaa aaaaaaatgg ttaaagttat
     2281 cggtcgtcgt tccttcggag tgcaaagaat atttgatatt ggtcttcccc aagaccataa
     2341 ttttctgcta gccaatgggg cgatcgccca caactgtcgt tgtgttttta gtggttgtct
     2401 cagtttcggc accgaaattt taaccgttga gtacggccca ttgtccattg gcaaaattgt
     2461 gagtgaagaa attaattgtt ctgtgtacag tgttgatcca aaagggaaag tttacaccca
     2521 ggcgatcgcc caatggcatg accggggaga gcaggaagta ttggaatatg aattggaaga
     2581 tggttcagta atccgagcta cctctgaccg ccgcttttta accaccgatt atcgactggt
     2641 ggcgatcgaa gaaatttttg caaggcaact ggacttgttg actttagaaa aaaattaaat
     2701 taacggcact gcgaaaaaac cccgccgaag cggggttttt tgcgaaatat aatgaccctc
     2761 ttgataaccc aagagggcat tttttaatgc ccatggcgta cggatcttag aaaaactcat
     2821 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     2881 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     2941 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     3001 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     3061 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     3121 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     3181 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     3241 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     3301 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     3361 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     3421 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     3481 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     3541 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     3601 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     3661 ttattgttca tgaagagaaa cctacggatc g
//
