LOCUS       40924_32963        5650 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transfection vector pN-PURO-PTP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5650)
  AUTHORS   Schimanski B, Nguyen TN, Gunzl A.
  TITLE     Highly efficient tandem affinity purification of trypanosome protein
            complexes based on a novel epitope combination
  JOURNAL   Eukaryotic Cell 4 (11), 1942-1950 (2005)
  PUBMED    16278461
REFERENCE   2  (bases 1 to 5650)
  AUTHORS   Schimanski B, Nguyen TN, Gunzl A.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-AUG-2005) Department of Genetics and Developmental 
            Biology, University of Connecticut Health Center, 263 Farmington 
            Ave, Farmington, CT 06030, USA
REFERENCE   3  (bases 1 to 5650)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5650)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Eukaryotic 
            Cell"; date: "2005"; volume: "4"; issue: "11"; pages: "1942-1950"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-AUG-2005) Department of Genetics and Developmental Biology, 
            University of Connecticut Health Center, 263 Farmington Ave, 
            Farmington, CT 06030, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /label=AmpR promoter
     CDS             126..983
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1157..1745
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2069..2099
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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)."
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2199..2204
                     /note="unique SacI restriction site for cloning the
                     resistance marker cassette"
     misc_feature    2204..2702
                     /label=HSP70 genes 2 and 3 intergenic region
                     /note="HSP70 genes 2 and 3 intergenic region"
     CDS             2703..3299
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    3315..4055
                     /label=beta-alpha tubulin intergenic region
                     /note="beta-alpha tubulin intergenic region"
     misc_feature    4056..4061
                     /note="unique HindIII restriction site for cloning the 
                     resistance marker cassette and the PTP cassette"
     CDS             4526..4699
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
     CDS             4703..4873
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             4910..4930
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    4973..4998
                     /note="multiple cloning site containing unique restriction
                     sites for KpnI, ApaI, SnaBI and NotI for fusing the target 
                     protein to the PTP sequence"
     promoter        complement(5007..5025)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5032..5048)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5190..5645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctcgaaat taaccctcac taaagggaac aaaagctgga gctccctaaa gtcgaggagg
     2221 ttgactaagt tcccaggtgt atttcggacc ggtgctgcag tcgtctcact gtctacggtg
     2281 atggtccgcc gccacatttt ctcttcgtct ctgtagcatt taggaaccct cgttgcggga
     2341 aagatgcgtg acacattgat gctactacta ttattaatac tctactatta ttattattat
     2401 tattattttt attttagttc cccaccaccg tcattattat tatcattatt attatcaata
     2461 ttaccattat tattattacg ataatgagcg ttagtgctat taccgctgtt gttgccatct
     2521 ttgttaccgt ggtgcttttt ttcctttggt tcatgcaact gcagcccggc atcgtcctcc
     2581 catacttgca ctcctgctct tccttcctat ttgcatttgc ccacgtgccc cacgcgtgca
     2641 tcacataagc atcaaggaac aagcaacagc tataaaacaa ggaatatctg cctctttgac
     2701 atatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     2761 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     2821 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     2881 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     2941 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     3001 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     3061 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     3121 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     3181 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     3241 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     3301 gatcgaactt cgaaaattca cggaggcaga gtccaacatg aacgatctcg tgtctgagta
     3361 ccagcagtac caggatgcca cgattgagga ggagggcgag ttcgacgagg aggagcaata
     3421 ctagacgcgg acggggcatt tcccgttcgt cattagcagt aggtaatgaa gatgtttgtt
     3481 tctcgtcccc tttctccttc gtccttctgt cattttgttc ttttgtgttt atgttttgtt
     3541 gttgttttct ttaatttttt ttttcttcca cgtttgtgta catccgcgcg ccactctatt
     3601 cagagagcca cggatagtag aggaggtggg aagggtatat gagggacacg cgtaccatga
     3661 tgtgggatgt attggggtcc ctgtctgtcc ttacgtgact atgtatgaac cgtcacgtgt
     3721 aagatgagct agtgagatca acagtacaac tcatcaacac gccttcttct cgttaaatgt
     3781 acacaatctt gatcctccac ctttatgggt cccattgttt gcctcttccg ctgtgtggag
     3841 tgcgcctaca cgcacttctc acttcgtaag tggtggtggc gtaagtattg cctaatgttg
     3901 actctatatt ctcctctcct caccccctcg cggtgctgat ttctgacaga tcttcaaaca
     3961 ctagtttaag caaaggacta ttcatccgtt tatattagca acagtaggta ctagcaccac
     4021 taacaacaac aacaaagcac ttctatttat ttatcaagct tctgaccttt gtaccatcgc
     4081 tgccctcacc tgttaactat ggtatttagt ggcctcagga ggttgcctgg gtggctgggt
     4141 ctttacttcg tcaggataat ttgaagttcg ttttccgtaa acagttaccc agttgtgagg
     4201 ttgttgcgac gcacattcac atgtcttcat cttatttgtt ggtatatttc cttactgctc
     4261 gttttggggt gcgggcgcgc tgattcaaag agaggggaga atctgccgcg ctgcttgcag
     4321 cacaagtctt gctgtttccc ccctctattg tcgtttttca cattcatgct atttccggtt
     4381 gaagcggtcg ttgtggccct gccgggtggt tctgcacatg ctatgcactt tctccctctt
     4441 cttcatcctc gtaatgctaa cagaataagt tgtggaaagc gtgaagcgtg caaacatggc
     4501 aggccttgcg caccacgatg aagccgtgga caacaaattc aacaaagaac aacaaaacgc
     4561 gttctatgag atcttacatt tacctaactt aaacgaagaa caacgaaacg ccttcatcca
     4621 aagtttaaaa gatgacccaa gccaaagcgc taacctttta gcagaagcta aaaagctaaa
     4681 tgatgctcag gcgccgaaag tagacaacaa attcaacaaa gaacaacaaa acgcgttcta
     4741 tgagatctta catttaccta acttaaacga agaacaacga aacgccttca tccaaagttt
     4801 aaaagatgac ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc taaatgatgc
     4861 tcaggcgccg aaagtagacg cgaattgtga tatacctaca actgcttctg aaaatttata
     4921 ttttcaaggt gaactagaag atcaggtgga tcctcgtctt attgatggga aagcggccgc
     4981 tacgtagggc ccggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt
     5041 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     5101 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5161 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     5221 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5281 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5341 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5401 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5461 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5521 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5581 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5641 aatttaggtg
//
