LOCUS       sequence-177974-        6759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for expression of N-His-3607 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6759)
  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 6759)
  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: Ppj184 (JBEI-13431)
FEATURES             Location/Qualifiers
     source          1..6759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     894..913
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(933..1010)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1216..1235
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1319..1338
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1319..1337
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1338..1362
                     /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             1377..1399
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1418..1435
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1445..1462
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2942..2959
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3012..3030)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3026..3073
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3110..3565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3197..3216)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3407..3428
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3657..4472)
                     /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(3657..4472)
                     /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     4380..4399
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4464..4483
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4594..5182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5083..5102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5336..5353
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5368..5507
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5433..5452)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5593..5615
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5609..5800)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6609..6759,1..932))
                     /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"
ORIGIN
        1 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
       61 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
      121 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
      181 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
      241 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
      301 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
      361 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
      421 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
      481 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
      541 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
      601 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
      661 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
      721 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
      781 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
      841 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
      901 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
      961 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     1021 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     1081 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     1141 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     1201 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     1261 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
     1321 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
     1381 tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc atcacagcag
     1441 cggcctggtg ccgcgcggca gccatatgag tagcaccaaa ccaattaaac cacatattca
     1501 aagtaccaag attgacacgc tgtatccgag ttacgatgtg attgttattg gtgccggtat
     1561 cgccggtctg atttgtgccg ccttcctggc caaaagtggt aagaaagccc tgctgattga
     1621 gcaacatttc attccaggtg gctactgtac cagctttaaa cgtaagggtt tcctgtttga
     1681 tgcggcggtt catcatattg gcggttgcgg taaatggagc gtggtgggtc gttgtttcaa
     1741 aatgctgggc atcgaaatgg acttttacca tctggaccca atggataatc tgatcttccc
     1801 aacgtttagc attgaaattc cagccgatct ggatgaatac ctgctgcgcc tgcaaaagcg
     1861 ttacccacat gaaaaggata acattaagtg gttttttcaa gactttatcc gcctgtaccg
     1921 cgccaccttc cataatgaga aaagtcaaat catcgagaaa tacaaagata tgacgtacgg
     1981 catgatgctg gatcatttct ttgatgatag tgaactgaaa atggttctga gcggccagtg
     2041 gggttacgtg ggtctggccc cgagcaaggc cagcgccatt gccatgagcc aaatgatgat
     2101 taactatctg aaagatggtg ccttttttcc aaagggtggt acccaagaat ttgccaatgc
     2161 catttttcgt aagtttatcg atttcggcgg tcatgttatg ctgagtagta aagtgaagaa
     2221 aatcctgaag aagggcccaa gtgttaatgg tgtgaaactg aatgacgata aggaaattca
     2281 tagcagtgtt gtggtgagca atatcgatgc gcgtcagacc ttttatcatc tgctgaacga
     2341 cggtaatgat gtggatgccg tttacctgaa aaacattgag ggtatgaaag aaagttgtag
     2401 tttttttctg ctgtacctgg gtatcggtaa aggcattgac ctgaatacgc tgaagcgcgg
     2461 cttctacctg agtggtagca atttcgacag catcgacagt gaatggatgt atattagtgt
     2521 gccaaccagt atttgtcaag cgctgagtcc acaaaataag caaattatca gtatcgtggt
     2581 gtacaagaaa gaagagttct acaaaaatgt gaccgattgg gaactgctga aacagaatat
     2641 tgtgagcagt acgctggagc gcctggaaag ttacctgccg gatattaaga aacatattga
     2701 gatcattgaa gttgccaccc caaaaaccct ggaacgctac accctgaaca cgaacggtgc
     2761 cgcgtatggt tgggccgtga gtgttgacca aatctgggaa cagcgtctga gcccaacgac
     2821 gccgattagt ggtctgtatc tggcgggtca ttggacgcgc ccaggtccag gtatctgcgc
     2881 cgtggttagt agtggttgga gtgtggccaa cctgattctg aaaaattggc gttaactcga
     2941 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3001 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3061 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     3121 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3181 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3241 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3301 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3361 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3421 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3481 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3541 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3601 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
     3661 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     3721 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     3781 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     3841 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     3901 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     3961 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     4021 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     4081 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     4141 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     4201 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     4261 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     4321 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     4381 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     4441 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     4501 gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4561 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4621 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4681 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4741 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4801 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4861 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4921 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4981 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5041 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5101 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5161 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5221 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5281 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5341 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     5401 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5461 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     5521 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5581 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5641 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     5701 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     5761 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     5821 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     5881 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     5941 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     6001 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     6061 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     6121 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     6181 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     6241 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
     6301 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     6361 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     6421 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     6481 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     6541 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     6601 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6661 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     6721 accagtgaga cgggcaacag ctgattgccc ttcaccgcc
//
