LOCUS       sequence-447452-        8297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses precursor peptide and lanthipeptide synthetase from 
            Chondromyces LanIV (FAST-RiPPs) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8297)
  AUTHORS   Ayikpoe RS, Shi C, Battiste AJ, Eslami SM, Ramesh S, Simon MA, 
            Bothwell IR, Lee H, Rice AJ, Ren H, Tian Q, Harris LA, Sarksian R, 
            Zhu L, Frerk AM, Precord TW, van der Donk WA, Mitchell DA, Zhao H
  TITLE     A scalable platform to discover antimicrobials of ribosomal origin.
  JOURNAL   Nat Commun. 2022 Oct 17;13(1):6135. doi: 10.1038/s41467-022-33890-w.
  PUBMED    36253467
REFERENCE   2  (bases 1 to 8297)
  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 
            Commun."; date: "2022-10-17"; volume: "13(1):6135. doi"; pages: " 
            10.1038/s41467-022-33890-w"
COMMENT     Sequence Label: pET28:LanIV-Chondromyces
FEATURES             Location/Qualifiers
     source          1..8297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2016..3098)
                     /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     3060..3079
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3099..3176)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3382..3401
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3489..3507
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3508..3532
                     /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)."
     CDS             3549..3566
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3877..3924
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3929..3947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3948..3972
                     /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)."
     terminator      6726..6773
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6814..7269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6901..6920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7111..7132
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7361..8176)
                     /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(7361..8176)
                     /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     8084..8103
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8168..8187
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     1321 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     1381 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     1441 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     1501 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     1561 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     1621 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg
     1681 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     1741 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     1801 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     1861 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     1921 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     1981 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2041 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2101 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     2161 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     2221 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     2281 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     2341 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     2401 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     2461 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     2521 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     2581 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     2641 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     2701 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     2761 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     2821 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     2881 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     2941 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     3001 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     3061 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     3121 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     3181 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     3241 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     3301 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     3361 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     3421 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     3481 aaatatggta atacgactca ctatagggga attgtgagcg gataacaatt ccaaggagat
     3541 atacaatgca ccaccaccac caccacgcaa tgaagaatca gtttggtgac gtccgcggtg
     3601 atatgacgga aattttggat caggcacgca aagatgctgc gttttctgag gaactgttaa
     3661 cgaacggtat gcctatggcg aaggaagaag atgagagcga cggggcaacg gtgcgcaaat
     3721 gtatgctgtt gacaattatt taaatatgtt gcgtttcttc gaccttttaa ccgctccctt
     3781 aggagaaaga cagatagctt cttacccgta ctccaccgtt ggcagcacga tcgcatgtcc
     3841 cacgtgaacc attggtaaac cctgtggcct gtcggtctag cataacccct tggggcctct
     3901 aaacgggtct tgaggggttt tttgccctta atacgactca ctatagggga attgtgagcg
     3961 gataacaatt ccaaggagat atacaatgaa tcccgaatct aatgcatctc ttcgttgcct
     4021 gcctggagca gatggtttct ttgacattat tcagagtgcg tgtttctcac cgcgtgcacg
     4081 cgcttccgga cgtcgtccac gtgctcctgc gggaccgtgg gatccgtgga tttgtgtgtt
     4141 gggagaagaa tatcctgcac ctgcccaagg ctggaaattg catttatccg cgacggttgg
     4201 ctcggctgaa gaaattctgc gccgtgcact tccgatcctg ctcgcagaag atgcagtctt
     4261 taaagttgca tcatcgacga caaccctggc ggctctgaac caaggtgaat ctggacttgg
     4321 tcaagtaggt aaatttatca cagtgtatcc ccgtgatgat gcacatgcag tgcgtttagc
     4381 ggcgcgtctt gatgaagcga cacgtggcct tcctggcccc acaattccta gcgatcgtgc
     4441 gttgcgccct ggctcccgtg ttttctatcg ttacggttca tttggcggac aagcactgca
     4501 attggccaca ggagaaattg ttgcggcaat gaaaacccca gatggtcaac ttgtgccaga
     4561 tcgtcgtcct gcagattacg acgtaccttc ttgggccaca gacccattta ttgcagcagg
     4621 cctggcagct gaattacctc gtggtgcttc acgtcatgta attgctcgtc gttatgttcc
     4681 tgcagcggtt ctgcaacgta ctccgcgtag taccatcttt ctggctctgg atttaaaagc
     4741 gtatcgtcgt tgtgtactta aacgtgtgga acgctgggat ggagcacgtt tgcgccatga
     4801 agcagatgta ttgcgtcgtc ttgggccaga tcccggtttt ccagcactgt atgacgtgtt
     4861 tgaagatgaa gaacacttat atttggtttt agaagatctg tctgctgaaa ctttatcggc
     4921 acatgttgca cacgccgcgg catctggttg ttttctgcca gcagcccaga ttgtgactct
     4981 gggccgcgcg ctggctcgtt tacttggtaa actgcacgca gagggactgg ctcatggtga
     5041 tctcaaaagc agtaatgtgc tggtggcacc tgcgaccctt cacctctggc ttattgactt
     5101 cgaactggcg catggcccat ctttgacggc acagccgcat cacgggcatg gtacccgtgg
     5161 ataccgtagc cctgagcgta ttgcaggtgc gccgcctagt acttcagatg acattcatgc
     5221 ctttggtgct ctcctgtact ttctggctac aggtgcggaa ccggccgagg gacctacaga
     5281 agccccgctg gatggtcgtc caccacgttt gatgaatcct aatttaccgc caggattgtg
     5341 cactgtgatt gctcgttcat tagcacagga tcctagcgca cgttatagta gcgtgcgcca
     5401 gattgaacgt gccttggaag atgttcttac tgacagtgcc acgcctttaa ccgttcccgc
     5461 tccaccgcgt catttcggcc gtgcgagtgc gcatttacct ttcccagccc gtcgtgaagc
     5521 acttgaactt gcccgccgtt taggcgattc gctggtcagc gctgctcgtc cggcacctcg
     5581 cggtggcggc gtattttggc cagatatgca tcctgacgcg cgcggacttc catctcgcga
     5641 tattggccat ggtacagcag gagtggttct ggcgttatcc tcgcttgtcg caacattggg
     5701 tgaatcaagt catcgcgcca ctttagcacg tgcggctttc ggactgctgc atgccccgcg
     5761 tccggatggg gctccggtgg ccggtttata cttgggcgaa gctggtattg gcgcggccct
     5821 tttaggtgct ggccgtgtgt tgggtgaccg tacacttgcg cgtgccgcag aggcacgcgg
     5881 tcgttggatc gccaccctgc cttttggatc accagatctt tataatggcg tagctggtcg
     5941 tttacgtttt cacctggcct tgcatcgtgc gacttcttct gacgaacatc tttcagcggc
     6001 tctccgtgca ggagatcacc tggttgatgt ggccgaatgc gatgaggatg ggaatgtgct
     6061 ttggcgcttt ccggaagggc atggcaactt aagtggtact actcaacttg gttatgccca
     6121 tggtgcagcc ggcattgcgg atagccttct ggatttattt gaagtcactc gtgaagttcg
     6181 ttacctggat gcagcacgtg gtgcggcccg ctggttggcc acacttgcag ttcccgcctt
     6241 gggcgatggt tcggggagcg cctggcctgt ggttcccggc ggtccgccac atgctgcaca
     6301 ctggtgtcat ggagccacag ggattggtcg tttcttctta catgcaaccg cattagaact
     6361 cgcacccgga tggacggatt tagccgcgcg tgcggcacgc accgtagccc gtggagcgcg
     6421 ctggttggga ccgactcagt gtcatggcct tgcaggtaat ttagaattcc ttgttgatat
     6481 gtttcaagca acgtcacacc cgcattattt gaccgaagcg gcggatttgt ttacccttct
     6541 gcgtgcattt agtcaagaaa aggcaggtcg tctggtctgg caaggcgatc aaccggaaca
     6601 ggctctgcct gcattgcagg tgggctatgg tggtgttgca acctgtttac ttcgtttggg
     6661 cgatccagct cgtgcacctc atttgctttc agtacgtggt ttaggtggtg ccggtgagta
     6721 acggtctagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgagcgctg
     6781 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     6841 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6901 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6961 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7021 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7081 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7141 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     7201 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     7261 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7321 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
     7381 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     7441 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     7501 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     7561 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     7621 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     7681 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     7741 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     7801 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     7861 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     7921 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     7981 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     8041 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     8101 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     8161 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     8221 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8281 aagatcaaag gatcttc
//
