LOCUS       pET-44b(+).        7310 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing 6xHis-tagged NusA fusion proteins. 
            For other reading frames, use pET-44a(+) or pET-44c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-44b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7310)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7310)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      complement(141..168)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(260..346)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(512..529)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(536..568)
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             complement(680..694)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(713..730)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(746..790)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(800..817)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(833..2317)
                     /label=NusA
                     /note="transcription elongation protein (N utilization
                     substance protein A)"
     CDS             complement(2324..2341)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(2351..2353)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(2361..2383)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2398..2422)
                     /label=lac operator
                     /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)."
     promoter        complement(2423..2441)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2754..2831
                     /label=lacI promoter
     CDS             2832..3911
                     /label=lacI
                     /note="lac repressor"
     protein_bind    3927..3948
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4486..4674
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4779..4921
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5107..5695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5776..5868
                     /label=AmpR promoter
     CDS             5869..6726
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(6844..7299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca
      121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
      181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
      241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
      301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
      361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
      421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
      481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
      541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
      601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
      661 gatccacgag ctccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc
      721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag
      781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt
      841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca
      901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa
      961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat
     1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg
     1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca
     1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca
     1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt
     1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac
     1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat
     1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa
     1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga
     1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac
     1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac
     1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca
     1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg
     1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga
     1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat
     1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt
     1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa
     1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa
     2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct
     2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc
     2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg
     2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg
     2281 atacggcttc aactacagcc aaaatttctt tgttcatact agaatggtga tggtggtgat
     2341 ggctgctgcc catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
     2401 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
     2461 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     2521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     2581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     2641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     2701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     2761 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     2821 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     2881 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     2941 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     3001 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     3061 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     3121 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     3181 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     3241 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     3301 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     3361 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     3421 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     3481 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     3541 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     3601 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
     3661 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     3721 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     3781 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3901 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     3961 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     4021 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     4081 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     4141 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     4201 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     4261 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     4321 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     4381 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     4441 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     4501 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     4561 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     4621 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     4801 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     4861 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5761 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5821 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa
     5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc
     6841 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     7021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     7081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     7141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     7201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     7261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
