LOCUS       sequence-332011-        7096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of His6-TEV-PRC1 in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7096)
  AUTHORS   Subramanian R, Ti SC, Tan L, Darst SA, Kapoor TM
  TITLE     Marking and measuring single microtubules by PRC1 and kinesin-4.
  JOURNAL   Cell. 2013 Jul 18;154(2):377-90. doi: 10.1016/j.cell.2013.06.021.
  PUBMED    23870126
REFERENCE   2  (bases 1 to 7096)
  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: 
            "10.1016/j.cell.2013.06"; journalName: "Cell"; date: "2013-07-18- 
            18"; volume: "154"; issue: "2"; pages: "377-90"
COMMENT     Sequence Label: His-TEV-PRC1
FEATURES             Location/Qualifiers
     source          1..7096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     380..399
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(419..496)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     702..721
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     809..828
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        809..827
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    828..852
                     /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             867..889
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             908..925
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             965..985
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2870..2914
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2952..2970)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2966..3013
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3050..3505
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3347..3368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3620..4411,4412..4480))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4243..4262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4481..4573)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4654..5242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5143..5162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5396..5413
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5428..5567
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5493..5512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5653..5675
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5669..5860)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6432..7096,1..418))
                     /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 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
       61 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
      121 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
      181 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
      241 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
      301 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
      361 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
      421 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
      481 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
      541 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
      601 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
      661 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
      721 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
      781 atcgagatcg atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      841 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
      901 cagcagccat caccatcatc accacagcca ggatccgaat tcggattacg atatcccaac
      961 gaccgaaaac ctgtattttc agggcgcggc cgcaatgagg agaagtgagg tgctggcgga
     1021 ggagtccata gtatgtctgc agaaagccct aaatcacctt cgggaaatat gggagctaat
     1081 tgggattcca gaggaccagc ggttacaaag aactgaggtg gtaaagaagc atatcaagga
     1141 actcctggat atgatgattg ctgaagagga aagcctgaag gaaagactca tcaaaagcat
     1201 atccgtctgt cagaaagagc tgaacactct gtgcagcgag ttacatgttg agccatttca
     1261 ggaagaagga gagacaacca tcttgcaact agaaaaagat ttgcgcaccc aagtggaatt
     1321 gatgcgaaaa cagaaaaagg agagaaaaca ggaactgaag ctacttcaag agcaagatca
     1381 agaactgtgc gaaattcttt gtatgcccca ctatgatatt gacagtgcct cagtgcccag
     1441 cttagaagag ctgaaccagt tcaggcaaca tgtgacaact ttgagggaaa caaaggcttc
     1501 taggcgtgag gagtttgtca gtataaagag acagatcata ctgtgtatgg aagaattaga
     1561 ccacacccca gacacaagct ttgaaagaga tgtggtgtgt gaagacgaag atgccttttg
     1621 tttgtctttg gagaatattg caacactaca aaagttgcta cggcagctgg aaatgcagaa
     1681 atcacaaaat gaagcagtgt gtgaggggct gcgtactcaa atccgagagc tctgggacag
     1741 gttgcaaata cctgaagaag aaagagaagc tgtggccacc attatgtctg ggtcaaaggc
     1801 caaggtccgg aaagcgctgc aattagaagt ggatcggttg gaagaactga aaatgcaaaa
     1861 catgaagaaa gtgattgagg caattcgagt ggagctggtt cagtactggg accagtgctt
     1921 ttatagccag gagcagagac aagcttttgc ccctttctgt gctgaggact acacagaaag
     1981 tctgctccag ctccacgatg ctgagattgt gcggttaaaa aactactatg aagttcacaa
     2041 ggaactcttt gaaggtgtcc agaagtggga agaaacctgg aggcttttct tagagtttga
     2101 gagaaaagct tcagatccaa atcgatttac aaaccgagga ggaaatcttc taaaagaaga
     2161 aaaacaacga gccaagctcc agaaaatgct gcccaagctg gaagaagagt tgaaggcacg
     2221 aattgaattg tgggaacagg aacattcaaa ggcatttatg gtgaatgggc agaaattcat
     2281 ggagtatgtg gcagaacaat gggagatgca tcgattggag aaagagagag ccaagcagga
     2341 aagacaactg aagaacaaaa aacagacaga gacagagatg ctgtatggca gcgctcctcg
     2401 aacacctagc aagcggcgag gactggctcc caatacaccg ggcaaagcac gtaagctgaa
     2461 cactaccacc atgtccaatg ctacggccaa tagtagcatt cggcctatct ttggagggac
     2521 agtctaccac tcccccgtgt ctcgacttcc tccttctggc agcaagccag tcgctgcttc
     2581 cacctgttca gggaagaaaa caccccgtac tggcaggcat ggagccaaca aggagaacct
     2641 ggagctcaac ggcagcatcc tgagtggtgg gtaccctggc tcggcccccc tccagcgcaa
     2701 cttcagcatt aattctgttg ccagcaccta ttctgagttt gcgaaggatc cgtccctctc
     2761 tgacagttcc actgttgggc ttcagcgaga actttcaaag gcttccaaat ctgatgctac
     2821 ttctggaatc ctcaattcaa ccaacatcca gtcctgactc gagtctggta aagaaaccgc
     2881 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
     2941 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3001 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     3061 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3121 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3181 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3241 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3301 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3361 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3421 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3481 aacaaaatat taacgtttac aatttctggc ggcacgatgg catgagatta tcaaaaagga
     3541 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3601 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3661 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3721 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3781 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3841 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3901 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3961 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4021 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4081 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4141 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4201 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4261 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4321 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4381 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4441 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatcat
     4501 gattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4561 aaaaataaac aaataggtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4621 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4681 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4741 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4801 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4861 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4921 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4981 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5041 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5101 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5161 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5221 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5281 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5341 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5401 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     5461 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5521 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     5581 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5641 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5701 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     5761 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     5821 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     5881 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     5941 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     6001 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     6061 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     6121 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     6181 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     6241 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     6301 acaggagcac gatcatgcta gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     6361 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     6421 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6481 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     6541 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     6601 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     6661 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca
     6721 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     6781 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     6841 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     6901 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     6961 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     7021 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     7081 gccggaacat tagtgc
//
