LOCUS       sequence-177983-        6739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for expression of N-StrepII-3607.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6739)
  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 6739)
  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: Ppj219(JBEI-13475)
FEATURES             Location/Qualifiers
     source          1..6739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     955..974
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(994..1071)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1277..1296
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1380..1399
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1380..1398
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1399..1423
                     /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             1438..1460
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1477..1500
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2983..3000
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3053..3071)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3067..3114
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3151..3606
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3238..3257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3448..3469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3698..4513)
                     /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(3698..4513)
                     /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     4421..4440
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4505..4524
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4635..5223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5124..5143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5377..5394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5409..5548
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5474..5493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5634..5656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5650..5841)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6650..6739,1..993))
                     /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 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
       61 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
      121 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
      181 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
      241 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
      301 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
      361 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
      421 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
      481 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
      541 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
      601 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
      661 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
      721 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
      781 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
      841 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
      901 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
      961 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     1021 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     1081 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     1141 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     1201 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     1261 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     1321 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     1381 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     1441 gtttaacttt aagaaggaga tatacatatg gcgtcgtggt ctcatccgca gtttgaaaaa
     1501 ggcgccatga gtagcaccaa accaattaaa ccacatattc aaagtaccaa gattgacacg
     1561 ctgtatccga gttacgatgt gattgttatt ggtgccggta tcgccggtct gatttgtgcc
     1621 gccttcctgg ccaaaagtgg taagaaagcc ctgctgattg agcaacattt cattccaggt
     1681 ggctactgta ccagctttaa acgtaagggt ttcctgtttg atgcggcggt tcatcatatt
     1741 ggcggttgcg gtaaatggag cgtggtgggt cgttgtttca aaatgctggg catcgaaatg
     1801 gacttttacc atctggaccc aatggataat ctgatcttcc caacgtttag cattgaaatt
     1861 ccagccgatc tggatgaata cctgctgcgc ctgcaaaagc gttacccaca tgaaaaggat
     1921 aacattaagt ggttttttca agactttatc cgcctgtacc gcgccacctt ccataatgag
     1981 aaaagtcaaa tcatcgagaa atacaaagat atgacgtacg gcatgatgct ggatcatttc
     2041 tttgatgata gtgaactgaa aatggttctg agcggccagt ggggttacgt gggtctggcc
     2101 ccgagcaagg ccagcgccat tgccatgagc caaatgatga ttaactatct gaaagatggt
     2161 gccttttttc caaagggtgg tacccaagaa tttgccaatg ccatttttcg taagtttatc
     2221 gatttcggcg gtcatgttat gctgagtagt aaagtgaaga aaatcctgaa gaagggccca
     2281 agtgttaatg gtgtgaaact gaatgacgat aaggaaattc atagcagtgt tgtggtgagc
     2341 aatatcgatg cgcgtcagac cttttatcat ctgctgaacg acggtaatga tgtggatgcc
     2401 gtttacctga aaaacattga gggtatgaaa gaaagttgta gtttttttct gctgtacctg
     2461 ggtatcggta aaggcattga cctgaatacg ctgaagcgcg gcttctacct gagtggtagc
     2521 aatttcgaca gcatcgacag tgaatggatg tatattagtg tgccaaccag tatttgtcaa
     2581 gcgctgagtc cacaaaataa gcaaattatc agtatcgtgg tgtacaagaa agaagagttc
     2641 tacaaaaatg tgaccgattg ggaactgctg aaacagaata ttgtgagcag tacgctggag
     2701 cgcctggaaa gttacctgcc ggatattaag aaacatattg agatcattga agttgccacc
     2761 ccaaaaaccc tggaacgcta caccctgaac acgaacggtg ccgcgtatgg ttgggccgtg
     2821 agtgttgacc aaatctggga acagcgtctg agcccaacga cgccgattag tggtctgtat
     2881 ctggcgggtc attggacgcg cccaggtcca ggtatctgcg ccgtggttag tagtggttgg
     2941 agtgtggcca acctgattct gaaaaattgg cgttaactcg agcaccacca ccaccaccac
     3001 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     3061 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     3121 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3181 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3241 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3301 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3361 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3421 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3481 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3541 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3601 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3661 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     3721 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3781 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     3841 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     3901 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     3961 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     4021 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     4081 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     4141 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     4201 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     4261 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     4321 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     4381 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     4441 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     4501 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     4561 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4621 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4681 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4741 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4801 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4861 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4921 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4981 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5041 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5101 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5161 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5221 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5281 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5341 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5401 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     5461 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     5521 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     5581 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5641 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     5701 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     5761 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5821 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     5881 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     5941 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     6001 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     6061 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     6121 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     6181 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     6241 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     6301 cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     6361 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     6421 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     6481 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     6541 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     6601 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     6661 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6721 aggcggtttg cgtattggg
//
