LOCUS       sequence-175165-        6688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression of K. stuttgartiensis protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6688)
  AUTHORS   Javidpour P, Deutsch S, Mutalik VK, Hillson NJ, Petzold CJ, Keasling
            JD, Beller HR
  TITLE     Investigation of Proposed Ladderane Biosynthetic Genes from Anammox 
            Bacteria by Heterologous Expression in E. coli.
  JOURNAL   PLoS One. 2016 Mar 14;11(3):e0151087. doi: 
            10.1371/journal.pone.0151087. eCollection 2016.
  PUBMED    26975050
REFERENCE   2  (bases 1 to 6688)
  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: "PLoS One.";
            date: "2016-03-14"; volume: "11(3):e0151087. doi"; pages: " 
            10.1371/journal.pone.0151087. eCollection 2016"
COMMENT     Sequence Label: JPUB_006834
FEATURES             Location/Qualifiers
     source          1..6688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(112..1194)
                     /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     1156..1175
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1195..1272)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1478..1497
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1581..1600
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1581..1599
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1600..1624
                     /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)."
     RBS             1639..1661
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3133..3150
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3203..3221)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3217..3264
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3301..3756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3388..3407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3598..3619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3848..4663)
                     /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(3848..4663)
                     /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     4571..4590
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4655..4674
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4785..5373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5274..5293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5527..5544
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5559..5698
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5624..5643)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5784..5806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5800..5991)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
       61 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
      121 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
      181 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
      241 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
      301 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
      361 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
      421 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
      481 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
      541 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
      601 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
      661 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
      721 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
      781 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
      841 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
      901 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
      961 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     1021 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     1081 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     1141 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     1201 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     1261 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     1321 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     1381 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     1441 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     1501 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     1561 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     1621 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat gaagaagatg
     1681 atgctgatta acccacataa accaggtcgt catggtgagg aaagcattac ggtgattgtg
     1741 caaatgccgc tgaatctggc ctacattaaa gccctgaccc caggcgattg ggaatttgat
     1801 gtgattgacg aaaacatcga actggccatt gatgataacg gtgaactgac cttcgcgcca
     1861 gtggatctgg tgtgtattac cagtgtgacc taccaaagtc cacgtgccta caagattgcc
     1921 accgcctgta aacgtaaagg tatgaccgtg atcatgggtg gtattcatgc gagcgtgatg
     1981 ccagaagaag ccagtaagta cgttgacacc gtgtttattg gtgaagccga ggaagtgtgg
     2041 ccgcgtgtta ttaaggattt cgaagccggt aaactgaaaa aagtttatga cggtggtctg
     2101 ccaccactga acctgatgaa acgtgtgttt ccagatcgtg aatttctgcg caagaaatac
     2161 aattacaaat ttagtagtat tgtgaccacc aagggttgtc caaattactg tgatttctgt
     2221 agcgtgccaa cctttcaagg ccgcaaattt cgtgaacgtc catacgaaga tgtgctggaa
     2281 gagctggcgg cgaccgacta caagggcctg atgctggccg aagataactt ctatggtcat
     2341 ggcaaacgta gtaacgaacg tgcccgtaac ctgtttaaag gcatggtgga acgtaatctg
     2401 caaaaagact ggctgggttt taccgccctg aacattagtc aagacaaaga aaccctggac
     2461 tacatggcca aaagtggttg ttttggtatg ctgatgggta tcgagagtac caatgaaacc
     2521 gttctggaga aaatgaacaa acaagttaat ctgaaactgg gtaccgaaag ctattatgac
     2581 tgtatccaaa agatccatga tgccggtctg gttacctggg gcagtgtggt gtttggtgcg
     2641 gatggtgatg gtaaagatag tttcaagcgc atgaccgatt ttatcctgga aaataacatt
     2701 gatattctga cgtttggcat taattgtcca tttccaaaaa cgcaactgta caaacgcctg
     2761 gatagtgaaa aacgtatttt tcgtaagaat tacccagatg actgggaata ttacgacacg
     2821 gcccatgtgg tgcatcgtct ggtggatatg acgctggagg actttattga gggtatgcaa
     2881 tacgtgtacg accatattta cgccggtgac aatctgcgta agcgttttcg caatagtatt
     2941 aagacgacga ataatccacg taacagtatg tttgcctttc gtgttggcag tgattggcaa
     3001 caagtgtttg accaagttct ggagaacctg cgcctgctgt atgatagtgg tgattactat
     3061 aaagattatt acaagagtaa tagcgtgagt gtgagtaaaa aaccgctggt tgaaagtatc
     3121 acgacgctcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
     3181 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     3241 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
     3301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3361 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3421 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3481 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3541 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3601 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3661 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3721 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     3781 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
     3841 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     3901 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     3961 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     4021 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     4081 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     4141 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     4201 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     4261 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     4321 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     4381 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     4441 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     4501 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     4561 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     4621 atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     4681 tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg
     4741 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4801 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4861 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4921 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4981 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5041 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5101 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5161 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5221 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5281 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5341 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5401 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5461 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5521 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     5581 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     5641 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     5701 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5761 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5821 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5881 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5941 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     6001 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     6061 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     6121 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     6181 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     6241 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     6301 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     6361 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     6421 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     6481 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     6541 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     6601 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     6661 gaagacagtc ataagtgcgg cgacgata
//
