LOCUS       sequence-289376-        5129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5129)
  AUTHORS   Diebold ML, Fribourg S, Koch M, Metzger T, Romier C
  TITLE     Deciphering correct strategies for multiprotein complex assembly by 
            co-expression: application to complexes as large as the histone 
            octamer.
  JOURNAL   J Struct Biol. 2011 Aug;175(2):178-88. doi: 
            10.1016/j.jsb.2011.02.001. Epub 2011 Feb 12.
  PUBMED    21320604
REFERENCE   2  (bases 1 to 5129)
  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.jsb.2011.02.001"; journalName: "J Struct Biol"; date: 
            "2011-08"; volume: "175"; issue: "2"; pages: "178-88"
COMMENT     Sequence Label: pnYC-NvGCvH_M
FEATURES             Location/Qualifiers
     source          1..5129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2294..2313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2402..2421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2402..2420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2421..2445
                     /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             2460..2482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2489..3142
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             3164..3184
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3215..3235
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3263..3280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3361..3379)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3375..3422
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3597..3602,3603..3619,3620..3625))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3737..4282
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4183..4202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4399..4416
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4808..4910
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4911..5129,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4977..4996)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
      541 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
      601 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt
      661 tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca
      721 gagcctgata aaaacggtta gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      781 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgcttg tttcggcgtg
      841 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ctgccggcac ctgtcctacg
      901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
      961 cggaaggagc taccggacag cggtgcggac tgttgtaact cagaataaga aatgaggccg
     1021 ctcatggcgt tgactctcag tcatagtatc gtggtatcac cggttggttc cactctctgt
     1081 tgcgggcaac ttcagcagca cgtaggggac ttccgcgttt ccagacttta cgaaacacgg
     1141 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     1201 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1261 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     1321 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     1381 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     1441 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1501 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     1561 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccagtga
     1621 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     1681 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     1741 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     1801 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1861 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1921 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1981 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     2041 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     2101 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     2161 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     2221 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2281 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2341 tgatgccggc cacgatgcgt ccggcgtaga ggatcagact aagatctcga tcccgcgaaa
     2401 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccact agtaataatt
     2461 ttgtttaact ttaagaagga gatataccat ggcccctata ctaggttatt ggaaaattaa
     2521 gggccttgtg caacccactc gacttctttt ggaatatctt gaagaaaaat atgaagagca
     2581 tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga
     2641 gtttcccaat cttccttatt atattgatgg tgatgttaaa ttaacacagt ctatggccat
     2701 catacgttat atagctgaca agcacaacat gttgggtggt tgtccaaaag agcgtgcaga
     2761 gatttcaatg cttgaaggag cggttttgga tattagatac ggtgtttcga gaattgcata
     2821 tagtaaagac tttgaaactc tcaaagttga ttttcttagc aagctacctg aaatgctgaa
     2881 aatgttcgaa gatcgtttat gtcataaaac atatttaaat ggtgatcatg taacccatcc
     2941 tgacttcatg ttgtatgacg ctcttgatgt tgttttatac atggacccaa tgtgcctgga
     3001 tgcgttccca aaattagttt gttttaaaaa acgtattgaa gctatcccac aaattgataa
     3061 gtacttgaaa tccagcaagt atatagcatg gcctttgcag ggctggcaag ccacgtttgg
     3121 tggtggcgac catccgccaa aatcggatgg taccggcagc ggcgaaaacc tttacttcca
     3181 gggccatatg ctcgagctta agcaattggg atccgaaaac ctttacttcc agggcagcgg
     3241 cgccatgggc agcggcagcg gccaccacca ccaccaccac ggtacctaat agtctagagc
     3301 tagccctagg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     3361 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3421 tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac
     3481 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     3541 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     3601 aagcttatcg atgataagct gtcaaacatg agaattacaa cttatatcgt atggggctga
     3661 cttcaggtgc tacatttgaa gagataaatt gcactgaaat caagaaatat tttatctgat
     3721 taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg
     3781 aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc
     3841 gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac
     3901 cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt
     3961 gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4021 tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa
     4081 ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa
     4141 ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct
     4201 ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc
     4261 aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt
     4321 atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc
     4381 agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac
     4441 atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga
     4501 caccctcatc agtgccaaca tagtaagcca gtatacactc ccctagcgct gatgtccggc
     4561 ggtgcttttg ccgttacgca ccaccccgtc agtagctgaa caggagggac agctgataga
     4621 aacagaagcc actggagcac ctcaaaaaca ccatcataca ctaaatcagt aagttggcag
     4681 catcacccga cgcactttgc gccgaataaa tacctgtgac ggaagatcac ttcgcagaat
     4741 aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat
     4801 gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata agatcactac
     4861 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     4921 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     4981 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     5041 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     5101 cccgcctgat gaatgctcat ccggaattc
//
