LOCUS       sequence-289496-        4442 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 4442)
  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 4442)
  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-NpS_N
FEATURES             Location/Qualifiers
     source          1..4442
                     /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..2524
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2546..2569
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2674..2692)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2688..2735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2910..2915,2916..2932,2933..2938))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3050..3595
                     /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     3496..3515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3712..3729
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4121..4223
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4224..4442,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4290..4309)
                     /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 gggcagcagc tggagccacc cgcagttcga
     2521 aaaaagcagc ggtaccggca gcggcctgga agttctgttc caggggcccc atatgctcga
     2581 gcttaagcaa ttgggatcct aatagtctag agctagccct aggagatccg gctgctaaca
     2641 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     2701 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     2761 atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt
     2821 gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg
     2881 ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac
     2941 atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt gaagagataa
     3001 attgcactga aatcaagaaa tattttatct gattaataag atgatcttct tgagatcgtt
     3061 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
     3121 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt
     3181 caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct
     3241 ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac
     3301 tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata
     3361 cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa
     3421 acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac
     3481 gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacca
     3541 ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg
     3601 ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc caggaaatct
     3661 ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca
     3721 gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac cggtgcagcc
     3781 ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca acatagtaag
     3841 ccagtataca ctcccctagc gctgatgtcc ggcggtgctt ttgccgttac gcaccacccc
     3901 gtcagtagct gaacaggagg gacagctgat agaaacagaa gccactggag cacctcaaaa
     3961 acaccatcat acactaaatc agtaagttgg cagcatcacc cgacgcactt tgcgccgaat
     4021 aaatacctgt gacggaagat cacttcgcag aataaataaa tcctggtgtc cctgttgata
     4081 ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt
     4141 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     4201 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     4261 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     4321 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     4381 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     4441 tc
//
