LOCUS       sequence-289494-        5930 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 5930)
  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 5930)
  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-NpHN_N
FEATURES             Location/Qualifiers
     source          1..5930
                     /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             2501..2518
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2534..4018
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization 
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="MDKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             4034..4057
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(4162..4180)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4176..4223
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4398..4403,4404..4420,4421..4426))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4538..5083
                     /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     4984..5003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5200..5217
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5609..5711
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5712..5930,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5778..5797)
                     /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 gagcggcagc catcatcatc atcatcacag
     2521 cggcagcggc tccatggaca aagaaatttt ggctgtagtt gaagccgtat ccaatgaaaa
     2581 ggcgctacct cgcgagaaga ttttcgaagc attggaaagc gccctggcga cagcaacaaa
     2641 gaaaaaatat gaacaagaga tcgacgtccg cgtacagatc gatcgcaaaa gcggtgattt
     2701 tgacactttc cgtcgctggt tagttgttga tgaagtcacc cagccgacca aggaaatcac
     2761 ccttgaagcc gcacgttatg aagatgaaag cctgaacctg ggcgattacg ttgaagatca
     2821 gattgagtct gttacctttg accgtatcac tacccagacg gcaaaacagg ttatcgtgca
     2881 gaaagtgcgt gaagccgaac gtgcgatggt ggttgatcag ttccgtgaac acgaaggtga
     2941 aatcatcacc ggcgtggtga aaaaagtaaa ccgcgacaac atctctctgg atctgggcaa
     3001 caacgctgaa gccgtgatcc tgcgcgaaga tatgctgccg cgtgaaaact tccgccctgg
     3061 cgaccgcgtt cgtggcgtgc tctattccgt tcgcccggaa gcgcgtggcg cccaactgtt
     3121 cgtcactcgt tccaagccgg aaatgctgat cgaactgttc cgtattgaag tgccagaaat
     3181 cggcgaagaa gtgattgaaa ttaaagcagc ggctcgcgat ccgggttctc gtgcgaaaat
     3241 cgcggtgaaa accaacgata aacgtatcga tccggtaggt gcttgcgtag gtatgcgtgg
     3301 cgcccgtgtt caggcggtgt ctactgaact gggtggcgag cgtatcgata tcgtcctgtg
     3361 ggatgataac ccggcgcagt tcgtgattaa cgcaatggca ccggcagacg ttgcttctat
     3421 cgtggtggat gaagataaac acacgatgga catcgccgtt gaagccggta atctggcgca
     3481 ggcgattggc cgtaacggtc agaacgtgcg tctggcttcg caactgagcg gttgggaact
     3541 caacgtgatg accgttgacg acctgcaagc taagcatcag gcggaagcgc acgcagcgat
     3601 cgacaccttc accaaatatc tcgacatcga cgaagacttc gcgactgttc tggtagaaga
     3661 aggcttctcg acgctggaag aattggccta tgtgccgatg aaagagctgt tggaaatcga
     3721 aggccttgat gagccgaccg ttgaagcact gcgcgagcgt gctaaaaatg cactggccac
     3781 cattgcacag gcccaggaag aaagcctcgg tgataacaaa ccggctgacg atctgctgaa
     3841 ccttgaaggg gtagatcgtg atttggcatt caaactggcc gcccgtggcg tttgtacgct
     3901 ggaagacctc gccgaacagg gcattgatga tctggctgat atcgaagggt tgaccgacga
     3961 aaaagccgga gcactgatta tggctgcccg taatatttgc tggttcggtg acgaagcggg
     4021 taccggcagc ggcctggaag ttctgttcca ggggccccat atgctcgagc ttaagcaatt
     4081 gggatcctaa tagtctagag ctagccctag gagatccggc tgctaacaaa gcccgaaagg
     4141 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     4201 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag
     4261 gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag
     4321 gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa
     4381 ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattaca
     4441 acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa
     4501 tcaagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
     4561 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     4621 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     4681 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     4741 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     4801 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     4861 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
     4921 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     5041 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     5101 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg
     5161 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     5221 gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg
     5281 ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact
     5341 cccctagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt cagtagctga
     5401 acaggaggga cagctgatag aaacagaagc cactggagca cctcaaaaac accatcatac
     5461 actaaatcag taagttggca gcatcacccg acgcactttg cgccgaataa atacctgtga
     5521 cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc gggaagccct
     5581 gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc aactttcacc
     5641 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     5701 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     5761 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     5821 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     5881 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
//
