LOCUS       40924_18011        7273 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing NusA fusion proteins. For other 
            reading frames, use pET-43.1a(+) or pET-43.1b(+).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7273)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7273)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7273
                     /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)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(536..568)
                     /codon_start=1
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
                     /translation="QPELAPEDPED"
     CDS             complement(679..693)
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     CDS             complement(712..729)
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             complement(745..789)
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(799..816)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(832..2316)
                     /codon_start=1
                     /label=NusA
                     /note="transcription elongation protein (N utilization
                     substance protein A)"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     RBS             complement(2324..2346)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2361..2385)
                     /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(2386..2404)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2717..2794
                     /label=lacI promoter
     CDS             2795..3874
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    3890..3911
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4449..4637
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    4742..4884
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5070..5658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5739..5831
                     /label=AmpR promoter
     CDS             5832..6689
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(6807..7262)
                     /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 ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg
      721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc
      781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc
      841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa
      901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac
      961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc
     1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc
     1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa
     1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag
     1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg
     1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc
     1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt
     1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac
     1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat
     1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc
     1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg
     1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac
     1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc
     1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa
     1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc
     1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg
     1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac
     1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac
     2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt
     2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct
     2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt
     2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga
     2281 tacggcttca actacagcca aaatttcttt gttcatatgt atatctcctt cttaaagtta
     2341 aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt
     2401 attaatttcg cgggatcgag atcgatctcg atcctctacg ccggacgcat cgtggccggc
     2461 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     2521 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     2581 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
     2641 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     2701 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     2761 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     2821 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     2881 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     2941 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3001 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     3061 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     3121 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     3181 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     3241 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     3301 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     3361 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     3421 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     3481 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     3541 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggacatctc
     3601 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     3661 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     3721 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     3781 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3841 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     3901 agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca
     3961 gctccttccg gtgggcgcgg ggcatgacta gcatgatcgt gctcctgtcg ttgaggaccc
     4021 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
     4081 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
     4141 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
     4201 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
     4261 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
     4321 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
     4381 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
     4441 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
     4501 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
     4561 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
     4621 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4681 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4741 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     4801 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc
     4861 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4921 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4981 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5041 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5101 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5161 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5221 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5281 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5341 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5401 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5461 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5521 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5581 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5641 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5701 aacgaaaact cacgttaagg gattttggtc atgacctatt tgtttatttt tctaaataca
     5761 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat catgattgaa
     5821 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5881 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5941 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6001 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6061 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6121 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6181 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6241 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6301 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6361 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6421 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6481 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6541 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6601 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6661 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6721 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6781 ataatctcat gccatcgtgc cgccagaaat tgtaaacgtt aatattttgt taaaattcgc
     6841 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     6901 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     6961 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     7021 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     7081 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     7141 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     7201 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     7261 gtcccattcg cca
//
