LOCUS       sequence-449503-        5190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses multinuclear non-heme iron dependent oxidative enzyme ChrH
            and partner protein ChrI in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5190)
  AUTHORS   Ayikpoe RS, Zhu L, Chen JY, Ting CP, van der Donk WA
  TITLE     Macrocyclization and Backbone Rearrangement During RiPP Biosynthesis
            by a SAM-Dependent Domain-of-Unknown-Function 692.
  JOURNAL   ACS Cent Sci. 2023 Apr 24;9(5):1008-1018. doi: 
            10.1021/acscentsci.3c00160. eCollection 2023 May 24.
  PUBMED    37252350
REFERENCE   2  (bases 1 to 5190)
  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: 
            "10.1021/acscentsci.3c00160"; journalName: "ACS Cent Sci"; date: 
            "2023-04-24- 24"; volume: "9"; issue: "5"; pages: "1008-1018"
COMMENT     Sequence Label: pCDFDuet-1:ChrH-ChrI
FEATURES             Location/Qualifiers
     source          1..5190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     promoter        1250..1268
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1269..1293
                     /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     complement(1857..1875)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1871..1918
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2089..2880)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2881..2972)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3020..3758)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3968..5050)
                     /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     5012..5031
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5051..5128)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5174..5190,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatataca tgaagaaacc tctccttgga ttagccatga tgccggaagc tgactttgtt
      121 tccgctattc ttccgctttt gcaaacgcaa agtgttgacg tactcgagtg gagcttcgat
      181 acgctgtatg acgtcgaaga gccggaatgg ctgtccggac tgcttgactt ctattcagac
      241 aattcacgcc tgttgggaca cggggtgtac tattcattat tcgacgcccg ctggatggag
      301 cgccaagaga tttggctcaa gaagcttaaa gaagaggtga agcgccgcaa gtacaatcat
      361 atcaccgagc attttggctt catgaacact gagaatttcc accaaggcgt acctcttcct
      421 gtaccacttc tgccgaagac gctgcaaatt gggaaggatc gcctgtctcg tctccaggac
      481 gccgtggaga tcccggtggg tgtcgagaat ctggcattct ctttcagcat ggatgacgtc
      541 aaggagcaag gtgaattctt ggaccgtttg gtggaagaca tcgacgggtt cctcatcctc
      601 gatctccaca atatttactg ccagagctgc aacttcaagg tggacatgat ggagatcatc
      661 aatttatatc cgctcgagaa ggtcaacgag atccacctct ccggcggctc ttggcaagag
      721 tcggcatacg gcaagaaacc tgtgcgccgt gatacgcatg acgaccgtat ccctgaagag
      781 attctgaatt tattacccga ggttttaatt cagtgccatc ctgagtacgt aattattgaa
      841 cgcttaggcc acacacttaa tacagaggtg gccaagcaaa ttttcttcga tgactttaac
      901 cgcgtaaaga aaatcttgga gttatcagat tacccagtcg gagaagagaa aatttggaat
      961 cgccagaata ccgtccactc gaagcctgtt gaagatctcc tcttgtatgc tgagcaaact
     1021 gaattaaccc gtttactttt cgatggtaat acgacggaat ctattaagaa tcgcgatttt
     1081 cattatttca agcctgaatc ctgggacgag gagatgatca caactgcgca acagatcatt
     1141 aagaagtgga acccgtaacg gcgcgcctgc aggtcgacaa gcttgcggcc gcataatgct
     1201 taagtcgaac agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc
     1261 actatagggg aattgtgagc ggataacaat tccccatctt agtatattag ttaagtataa
     1321 gaaggagata tacatatggc agatctcatg aaaatgacta ctgtatttct gctgattacc
     1381 gcggttttaa cagcattgat cgctgggctg ttctacgctt actcatgctc agtggtactt
     1441 ggcttaggca agttaagtga cacggagtat cttaaaagca tgcaaaatat caatcgtgaa
     1501 atcttgaacc ctgttttctt catgagcttc atgggtaccg ccgttctctt accagttgcg
     1561 acgtttcttt tccgtggtga gcaaccttcg ttcttattct tactgttagc ctcggccgcg
     1621 tacctgattg gcgtgtttgg tgttacgatc gtaggcaacg tcccgttaaa tgatatgttg
     1681 gacaaatttg atatctccgg ttcaaccatg gatgtagtca agcaaatgcg cgaccgtttc
     1741 gagaaccgct ggaatctcct gaataacgtg cgtactgtgt tttcagtaat ttctatcgca
     1801 ttagtggttt gcgcgtgtat ttggaaccgc cagctgacgt aactaggctg ctgccaccgc
     1861 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1921 gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt
     1981 aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc
     2041 atcgtggccg gatcttgcgg cccctcggct tgaacgaatt gttagacatt atttgccgac
     2101 taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga
     2161 ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg
     2221 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     2281 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc
     2341 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     2401 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     2461 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     2521 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     2581 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     2641 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     2701 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     2761 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     2821 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     2881 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2941 catatttgaa tgtatttaga aaaataaaca aatagctagc tcactcggtc gctacgctcc
     3001 gggcgtgaga ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga
     3061 taagcagggg actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat
     3121 aggctccgcc ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga
     3181 gccgtgaggc tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc
     3241 cccaccgttt ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac
     3301 cggatacctg ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac
     3361 actggtatct cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc
     3421 ccgttcagcc cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa
     3481 agcacggtaa aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt
     3541 ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac
     3601 agatttggtt gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga
     3661 cttaaccttc gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta
     3721 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga
     3781 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     3841 tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     3901 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     3961 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4021 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4081 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4141 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4201 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
     4261 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     4321 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     4381 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     4441 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     4501 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     4561 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     4621 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     4681 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     4741 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     4801 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     4861 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     4921 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     4981 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     5041 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     5101 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     5161 tgcattagga aattaatacg actcactata
//
