LOCUS       sequence-177973-        6693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for expression of C-His-3607.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6693)
  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 6693)
  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: Ppj187 (JBEI-13437)
FEATURES             Location/Qualifiers
     source          1..6693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1396..1587
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1581..1603)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1689..1828
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1744..1763
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1843..1860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2014..2602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2094..2113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2713..2732)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2724..3539
                     /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             2724..3539
                     /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     complement(2797..2816)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3631..4086)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3768..3789)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3980..3999
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4123..4170)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4166..4184
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4237..4254)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5731..5753
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5768..5792
                     /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(5792..5811)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5793..5811)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5895..5914)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6120..6197
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6198..6693,1..587)
                     /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     complement(6217..6236)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
       61 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
      121 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
      181 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
      241 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
      301 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
      361 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
      421 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
      481 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
      541 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      601 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
      661 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
      721 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
      781 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
      841 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
      901 caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg
      961 aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc
     1021 gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg
     1081 gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt
     1141 atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta
     1201 ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc
     1261 gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt
     1321 atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct
     1381 tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc
     1441 agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca
     1501 gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt
     1561 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     1621 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     1681 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
     1741 atgcggcatc agagcagatt gtactgagag tgcaccattg cggtgtgaaa taccgcacag
     1801 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     1861 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1921 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     2461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg tctgcttaca
     2701 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctctaggc
     2761 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     2821 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
     2881 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     2941 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     3001 atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat
     3061 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     3121 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     3181 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     3241 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     3301 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     3361 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     3421 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     3481 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaag
     3541 aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3601 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     3661 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3721 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3781 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3841 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3901 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3961 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4021 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4081 gcgcgtccca ttcgccaatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac
     4141 ccgtttagag gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc
     4201 agcttccttt cgggctttgt tagcagccgg atctcagtgg tggtggtggt ggtgctcacg
     4261 ccaatttttc agaatcaggt tggccacact ccaaccacta ctaaccacgg cgcagatacc
     4321 tggacctggg cgcgtccaat gacccgccag atacagacca ctaatcggcg tcgttgggct
     4381 cagacgctgt tcccagattt ggtcaacact cacggcccaa ccatacgcgg caccgttcgt
     4441 gttcagggtg tagcgttcca gggtttttgg ggtggcaact tcaatgatct caatatgttt
     4501 cttaatatcc ggcaggtaac tttccaggcg ctccagcgta ctgctcacaa tattctgttt
     4561 cagcagttcc caatcggtca catttttgta gaactcttct ttcttgtaca ccacgatact
     4621 gataatttgc ttattttgtg gactcagcgc ttgacaaata ctggttggca cactaatata
     4681 catccattca ctgtcgatgc tgtcgaaatt gctaccactc aggtagaagc cgcgcttcag
     4741 cgtattcagg tcaatgcctt taccgatacc caggtacagc agaaaaaaac tacaactttc
     4801 tttcataccc tcaatgtttt tcaggtaaac ggcatccaca tcattaccgt cgttcagcag
     4861 atgataaaag gtctgacgcg catcgatatt gctcaccaca acactgctat gaatttcctt
     4921 atcgtcattc agtttcacac cattaacact tgggcccttc ttcaggattt tcttcacttt
     4981 actactcagc ataacatgac cgccgaaatc gataaactta cgaaaaatgg cattggcaaa
     5041 ttcttgggta ccaccctttg gaaaaaaggc accatctttc agatagttaa tcatcatttg
     5101 gctcatggca atggcgctgg ccttgctcgg ggccagaccc acgtaacccc actggccgct
     5161 cagaaccatt ttcagttcac tatcatcaaa gaaatgatcc agcatcatgc cgtacgtcat
     5221 atctttgtat ttctcgatga tttgactttt ctcattatgg aaggtggcgc ggtacaggcg
     5281 gataaagtct tgaaaaaacc acttaatgtt atccttttca tgtgggtaac gcttttgcag
     5341 gcgcagcagg tattcatcca gatcggctgg aatttcaatg ctaaacgttg ggaagatcag
     5401 attatccatt gggtccagat ggtaaaagtc catttcgatg cccagcattt tgaaacaacg
     5461 acccaccacg ctccatttac cgcaaccgcc aatatgatga accgccgcat caaacaggaa
     5521 acccttacgt ttaaagctgg tacagtagcc acctggaatg aaatgttgct caatcagcag
     5581 ggctttctta ccacttttgg ccaggaaggc ggcacaaatc agaccggcga taccggcacc
     5641 aataacaatc acatcgtaac tcggatacag cgtgtcaatc ttggtacttt gaatatgtgg
     5701 tttaattggt ttggtgctac tcattgtata tctccttctt aaagttaaac aaaattattt
     5761 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
     5821 gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     5881 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     5941 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     6001 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     6061 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg
     6121 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     6181 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     6241 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     6301 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     6361 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     6421 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     6481 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     6541 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     6601 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     6661 ccatcaacag tattattttc tcccatgaag acg
//
