LOCUS       sequence-183019-        7955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of HisMBP-TZAPZNF9-11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7955)
  AUTHORS   Li JS, Miralles Fuste J, Simavorian T, Bartocci C, Tsai J, Karlseder
            J, Lazzerini Denchi E
  TITLE     TZAP: A telomere-associated protein involved in telomere length 
            control.
  JOURNAL   Science. 2017 Jan 12. pii: aah6752. doi: 10.1126/science.aah6752.
  PUBMED    28082411
REFERENCE   2  (bases 1 to 7955)
  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: "Science. 
            2017 Jan 12. pii: aah6752. doi: 10.1126/science.aah6752."
COMMENT     Sequence Label: sv272-HisMBP-TZAPZNF9-11
FEATURES             Location/Qualifiers
     source          1..7955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1272..1463
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1457..1479)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1565..1704
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1620..1639
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1719..1736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1890..2478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1970..1989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2589..2608)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2600..3415
                     /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             2600..3415
                     /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(2673..2692)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3507..3962)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3644..3665)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3856..3875
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3999..4046)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4042..4060
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4113..4130)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4137..4169)
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             complement(5634..5654)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(5709..6809)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(5712..5735)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             complement(6819..6836)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             6856..6878
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6893..6917
                     /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(6917..6936)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6918..6936)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7258..7335
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(7336..7955,1..463)
                     /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 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
       61 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
      121 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
      181 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
      241 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
      301 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
      361 aaccaccctt gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      421 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      481 taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc
      541 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
      601 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
      661 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
      721 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
      781 ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga
      841 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
      901 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
      961 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     1021 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     1081 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     1141 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     1201 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     1261 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     1321 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     1381 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     1441 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     1501 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1561 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     1621 ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc
     1681 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1741 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1801 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1861 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1921 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1981 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2041 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2101 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2161 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2221 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2281 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2341 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2401 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2461 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2521 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
     2581 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
     2641 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2701 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     2761 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2821 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2881 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     2941 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3001 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3061 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3121 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3181 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3241 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3301 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3361 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
     3421 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3481 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     3541 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3601 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3661 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3721 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3781 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3841 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3901 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3961 gtcccattcg ccaatccgga tatagttcct cctttcagca aaaaacccct caagacccgt
     4021 ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct
     4081 tcctttcggg ctttgttagc agccggatct cagtggtggt ggtggtggtg ctcgacatcc
     4141 tcggggtctt ccggggcgag ttctggctgg ctagcccgtt tgatctcgag tgcggccgca
     4201 agcttgtcga cggagctcga attcggggat ccctatgtgt cacagtcctc ggggacagca
     4261 gctgtgatga tgagggaggg ctccaggaca cctgggccgg tgaagctctc ctctgcacac
     4321 agtctctggc cggggagctg ggaggccagg ccgccctggg ccagggactc cacaatgacc
     4381 tccgccgagc ccacctccag ctctgcaggc ggctgcagcg ccaccaccac catcttctcg
     4441 tcctcgatga tcaggttgcg gagcttgcgc tgccgcgggt tgtagttctc tacccggtcg
     4501 tggatctcca tgtgcctccg caggtgggcc tgtcgggtaa acttgtagcc acactcggtg
     4561 cactcaaact tcctcacccc cttgtgccgt ctgacgtgca cacgcagttg ctccacggct
     4621 ttgaaggtct tgccgcatat ctggcagaag tggggccggc cctcctgatg gcggctggcc
     4681 acgtgcctca ggaggggccc cttctcagtg aagcgctgtt cacagaactc gcaactgaag
     4741 ggcctttccc cggtttcgac cggcaccgct gtaccttttc tgttctcagc aggctcagct
     4801 gctagggagc ccgcgctcag ggctggctca gctgcacagg gctttcggat tacatttgac
     4861 ttcctcctgg tcagcacagc ctcaggctca gacatgtaat cgccatcccc atcatcctcc
     4921 acttgaacca ccacttccca ctcattactg ccgccctgca gctgggcacc ttcagcctct
     4981 aagggccttg ggggcacttt gcagtcctcg ggtttcttgt cctcaagggg acaaggcttc
     5041 aaggcctgct tcagtttccc acagagggag gaggggccct cactctgagc tggggaatgg
     5101 agctggggcc tctgaggact atgactgcct ctgggctcct gatccctggg gactagaccc
     5161 agagtcctcg aaacttcctc ttcttccaag cctgccggct ccttgacgtg gctgttcaca
     5221 ttctgggagg cagggggccc cagcccactc tggccacctg ctgcctgtcc cactgaagtt
     5281 ttgggcttga agctctggca cagctctacg gcctctggca ctcgcaactc cctggctgcc
     5341 aggagcacct gatcccggtt ccctgaggtg agagcgaggt gaccagtgta gaaaaagtcc
     5401 aacaagaggc caaagatctc agcgaagcca gcagggagga cgacactgcc ccctgagcca
     5461 tccccgtaga ggctctggaa aaagtgactg cagcaggcaa ggacactcca gtgtgcctta
     5521 aacaccaggc cccccacgtc cagagtggcg tcgcagtact ggcccttctc ccgctgcttg
     5581 ttgagctcct gcagaaccct cacactgtgc tggacgaagg agccgtccat ggcgccctga
     5641 aaatacaggt tttcctcgat cccgaggttg ttgttattgt tattgttgtt gttgttcgag
     5701 ctcgaattag tctgcgcgtc tttcagggct tcatcgacag tctgacgacc gctggcggcg
     5761 ttgatcaccg cagtacgcac ggcataccag aaagcggaca tctgcgggat gttcggcatg
     5821 atttcacctt tctgggcgtt ttccatagtg gcggcaatac gtggatcttt cgccaactct
     5881 tcctcgtaag acttcagcgc tacggcaccc agcggtttgt ctttattaac cgcttccaga
     5941 ccttcatcag tcagcagata gttttcgagg aactcttttg ccagctcttt gttcggactg
     6001 gcggcgttaa tacctgcgct cagcacgcca acgaacggtt tggatggttg acccttgaag
     6061 gtcggcagta ccgttacacc ataattcact ttgctggtgt cgatgttgga ccatgcccac
     6121 gggccgttga tggtcatcgc tgtttcgcct ttattaaagg cagcttctgc gatggagtaa
     6181 tcggtgtctg cattcatgtg tttgttttta atcaggtcaa ccaggaaggt cagacccgct
     6241 ttcgcgccag cgttatccac gcccacgtct ttaatgtcgt acttgccgtt ttcatacttg
     6301 aacgcataac ccccgtcagc agcaatcagc ggccaggtga agtacggttc ttgcaggttg
     6361 aacatcagcg cgctcttacc tttcgctttc agttctttat ccagcgccgg gatctcttcc
     6421 caggtttttg gcgggttcgg cagcagatct ttgttataaa tcagcgataa cgcttcaaca
     6481 gcgatcgggt aagcaatcag cttgccgttg taacgtacgg catcccaggt aaacggatac
     6541 agcttgtcct ggaacgcttt gtccggggtg atttcagcca acaggccaga ttgagcgtag
     6601 ccaccaaagc ggtcgtgtgc ccagaagata atgtcagggc catcgccagt tgccgcaacc
     6661 tgtgggaatt tctcttccag tttatccgga tgctcaacgg tgactttaat tccggtatct
     6721 ttctcgaatt tcttaccgac ttcagcgaga ccgttatagc ctttatcgcc gttaatccag
     6781 attaccagtt taccttcttc gattttcata gaagaaccat ggtgatggtg atggtgagaa
     6841 gaacccatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt
     6901 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatctgtga
     6961 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     7021 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     7081 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     7141 tgctgattgg cgttgcgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
     7201 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
     7261 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     7321 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     7381 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     7441 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     7501 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     7561 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     7621 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     7681 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     7741 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     7801 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     7861 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     7921 ctgcgtctgg ctggctggca taaatatctc actcg
//
