LOCUS       sequence-204377-        6880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of an N-terminally truncated form of Drosophila 
            topoisomerase I with a C-terminal 6His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6880)
  AUTHORS   Khuong MT, Fei J, Cruz-Becerra G, Kadonaga JT
  TITLE     A simple and versatile system for the ATP-dependent assembly of 
            chromatin.
  JOURNAL   J Biol Chem. 2017 Nov 24;292(47):19478-19490. doi: 
            10.1074/jbc.M117.815365. Epub 2017 Oct 5.
  PUBMED    28982979
REFERENCE   2  (bases 1 to 6880)
  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: 
            "10.1074/jbc.M117.815365"; journalName: "J Biol Chem"; date: 
            "2017-11-24- 24"; volume: "292"; issue: "47"; pages: "19478-19490"
COMMENT     Sequence Label: pET-NDH6
FEATURES             Location/Qualifiers
     source          1..6880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     965..984
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1004..1081)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1287..1306
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1390..1409
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1390..1408
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1409..1433
                     /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             1448..1470
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3148..3165
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3218..3236)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3232..3279
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3316..3769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3401..3420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3611..3632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3861..4676)
                     /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(3861..4676)
                     /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     4584..4603
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4668..4687
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5275..5294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5528..5545
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5560..5699
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5625..5644)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5785..5807
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5801..5992)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6801..6880,1..1003))
                     /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 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
       61 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
      121 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
      181 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
      241 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
      301 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
      361 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
      421 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
      481 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
      541 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
      601 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
      661 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
      721 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
      781 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
      841 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
      901 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
      961 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     1021 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     1081 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     1141 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     1201 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     1261 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     1321 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     1381 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     1441 aaataatttt gtttaacttt aagaaggaga tataccatgg cgatatcgga tccggaggtt
     1501 tggcgatggt gggaggagga gaagcgtgcc gatggcgtca aatggtcaac gctggagcac
     1561 aagggacccg tgtttgcacc gcgatacgaa cgggtgccac gcaatgtacg gttctactac
     1621 gatggcaagc cgttggagct ctctgaggaa acggaggagg cagccacgtt ctatgccaag
     1681 atgctgaacc acgactactg caccaaggaa gtgttcaata acaacttctt caaggacttc
     1741 cgcaagtcca tgacgcccag agagagggag attatcaagg atttccgcaa gtgcaacttc
     1801 caggagatgt tcaactactt ccaagcggag tccgaaaagc gcaaggcagc cagcaaggag
     1861 gagaagctga tcaagaagaa cgaaaacgag gcgctgatga aggaattcgg attctgcatg
     1921 attgatgggc acaaggagaa gatcggtaac ttccgcctgg agccgccagg cctgttccgc
     1981 ggtcgtggcg agcatcccaa aatgggcatg atcaagcggc gcatccaagc cagcgatgtg
     2041 tccattaact gtggcaaaga ttcaaaggtg ccttcaccgc cgcccggatc tcgctggaag
     2101 gaggtgcgcc acgacaacac ggtcacctgg ctggcctcct ggatcgagaa tgtgcaggga
     2161 caggtcaagt acatcatgtt gaatccctcc tcgaaactca agggcgagaa ggatcacatt
     2221 aagtacgaga cggcgcgacg cctggacaag gtaatcgata agatacgtgc cacctatcgc
     2281 gacgagtgga agtccaagga gatgagggtt cgccagcgag cggtggccct ttacttcatc
     2341 gacaaactgg cgctgagagc aggcaacgaa aaggacgagg atcaggccga taccgtgggc
     2401 tgttgctcgc tacgcgtgga gcatgtccag ctccacaagg aactgaacgg aaaggagaac
     2461 gtggtggtct ttgatttccc cggtaaggac tccattcggt attacaacga ggtcgaggtg
     2521 gagaagcgtg tcttcaagaa cctcgaactg ttcatggagc acaagaagga gggcgacgac
     2581 ctcttcgatc gtctcaacac gcaggtgctt aatgagcatc tcaaggagct gatggaaggt
     2641 ctcacggcca aggtattccg tacgtacaac gcatcgaaaa cactgcaaag tcagctggat
     2701 ctgttgaccg atccgagcgc cacggtgccg gaaaagctgt tggcctacaa tcgcgccaac
     2761 cgtgccgtgg ccatcctctg taaccatcag cgttccgtgc ccaagagcca cgagaagtca
     2821 atggagaatc tgaaggagaa gatcaaggcc aagcgggaag ccatcgaaaa atgcgagtcc
     2881 gagtatcatt cgagagacga gaagaagggc aaacagctgg agcgcctaag ggatcagctg
     2941 aagaaactag aactacaaga gactgataga gacgagaata agaccattgc cctgggcaca
     3001 tctaagctaa actatcttga tccacgcata tcggtggcat ggtgtaaaaa gcatgacgtg
     3061 ccgatcgaga agatattcaa caaaacgcaa agaaccaaat ttctgtgggc cgttcatagc
     3121 gccgatgaga attatcgttt cctcgagcac caccaccacc accactgaga tccggctgct
     3181 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     3241 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     3301 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt ggtggttacg
     3361 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3421 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3481 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3541 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3601 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3661 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3721 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     3781 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3841 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     3901 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     3961 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     4021 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     4081 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     4141 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     4201 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     4261 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     4321 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     4381 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     4441 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     4501 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     4561 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     4621 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
     4681 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
     4741 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4801 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4861 ggtggtttgt ttgccggatc aagagctacc gaaggtaact ggcttcagca gagcgcagat
     4921 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4981 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5041 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5101 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5161 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5221 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5281 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5341 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cttttttacg
     5401 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5461 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5521 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     5581 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     5641 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     5701 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5761 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5821 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     5881 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     5941 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     6001 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     6061 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     6121 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     6181 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     6241 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     6301 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     6361 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     6421 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     6481 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     6541 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     6601 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     6661 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     6721 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     6781 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6841 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
//
