LOCUS       40924_41126        6781 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Episomal vector pSpiro-NptII-ESP-C-SF-TAP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6781)
  AUTHORS   Jerlstrom-Hultqvist J, Einarsson E, Svard SG.
  TITLE     Stable Transfection of the Diplomonad Parasite Spironucleus 
            salmonicida
  JOURNAL   Eukaryotic Cell 11 (11), 1353-1361 (2012)
  PUBMED    22983987
REFERENCE   2  (bases 1 to 6781)
  AUTHORS   Jerlstroem-Hultqvist JSE., Einarsson EM, Svaerd SG.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2012) Department of Cell and Molecular Biology, 
            Uppsala University, BMC, Box 596, Uppsala 751 24, Sweden
REFERENCE   3  (bases 1 to 6781)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6781)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Eukaryotic 
            Cell"; date: "2012"; volume: "11"; issue: "11"; pages: "1353-1361"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-MAR-2012) Department of Cell and Molecular Biology, Uppsala 
            University, BMC, Box 596, Uppsala 751 24, Sweden"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /label=AmpR promoter
     CDS             129..986
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1160..1748
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     3'UTR           complement(2214..2593)
                     /note="fructose bisphosphate aldolase from Spironucleus 
                     salmonicida"
     CDS             complement(2594..3385)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     5'UTR           complement(3386..3678)
                     /note="fructose bisphosphate aldolase from Spironucleus 
                     salmonicida"
     5'UTR           3738..4398
                     /note="encystation-specific protease-like protein from 
                     Spironucleus salmonicida"
     CDS             5935..5958
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             5995..6018
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             6034..6057
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     3'UTR           6064..6123
                     /note="ribosomal protein S15A from Spironucleus
                     salmonicida"
     promoter        complement(6138..6156)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6166..6182)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6324..6779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct gggtaccact agtcttctag
     2221 caatattgat aatagacacg ccaatgatag aattaacagg ttaacataca aattagtaaa
     2281 gaggagtata ccttatttat taacgaaata aaatcataga taagtaagta tataataatt
     2341 ttaaaagctg attaattcta gagataacgt tactttaata taatatttag taaaacctta
     2401 ttattaatat gaaacttgta ttcagtgaat taaaaaatca acaaacaact taaataagag
     2461 gtagattaca taaaactaat tcatttagtg ataacactat aactaagtac tctaaggagt
     2521 tttagccttc aaacttctaa aaattactat aagatatgta taataaaatt aatagaataa
     2581 agagtcaact tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     2641 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     2701 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     2761 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     2821 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     2881 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     2941 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     3001 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3061 gagcaaggtg ggatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3121 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     3181 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     3241 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3301 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     3361 cgtgcaatcc atcttgttca atcattactt gaggaagtgg tgataagatt taataacttt
     3421 aatctctatt tatgatataa taataatttt atatgtattt tgtttttgtg gtttatagag
     3481 taactgatgg tttgttctca atattatata tgatttgtga ataaccaaac acatcttgat
     3541 ttaacgtcaa acatatacaa aacctaatca atcatttatt cttaatcttt tataaccgat
     3601 ctttgagcaa tttcatatcc ctcataactc tagtaatctc ttctttgatt ttacccattc
     3661 tgagtgtctt gtctggaccc atatgaagct tgcttaatta agcgctagcg atatcggatc
     3721 ctgcagtcta gacgcgtgaa ctctcagttg ttatgagatt atgtcagttt cagacgttgg
     3781 ttgattgttg tgatagtgat gagtttgtat gataatcgga atgctaagta taaaatttat
     3841 cattatcaca gctaagtgtt aatatagata tgaatatttt aacctttaca agttgataag
     3901 atacaacata tatatttaag atttgattat aaaagtaagt ccgaaatata atattaagtg
     3961 aaatgtaact tattttcctg ttacctacca atttatacat ttatatttta tttttgtcta
     4021 gagcttaaaa tgtcaatcga tttataataa ttctattcat tatgatgtaa ttactattat
     4081 tgaataaaat tataaattat aaaaccagga aactacatta agtaattgtg aacgataaaa
     4141 cggatgtttg acatcaataa aatcgaattt gtatctacaa aggtgataat aatataaaaa
     4201 tataatatag agtcttgaaa tcacttaatt gtgaaccgta tttggtagta aagatcatta
     4261 ttgtttgata ttacttaata tttgtaaaaa caattctaac tctattctta agttaattaa
     4321 ataaatcgta atattttgag aaactttgag tagtataata tagataatga attacttaat
     4381 ccaaaattta atgcattgat ggctttatat attaatatta tttatttacg ttccacagat
     4441 ttagtaatgt tgcttttgtt gattgcagat acaccagcaa tgtgttacaa tgactaagtc
     4501 ttaggaacgt ggaatatcag tatttctaaa tatgattatt atcctaagaa tgataaagta
     4561 gtatgttccc ctaattttta ggttgacgag tataaagaaa taaccttaat tgcaccaaat
     4621 gttgccatag ataaaaaagg tattacaggt acctggacta tgatttatac tcaagctgtt
     4681 gagattaata ttgggaacaa taaatattat ttctaacttg actataagag agcgtcgtct
     4741 gaaagtgacc cggttcaatg gattgattct tactgtaatt agtctgaccc taacgccgcc
     4801 tgggcagttg aataaggtgt aacaagaggt gtttatgctt gtttcaccgc ggttaacatt
     4861 aatcctacca cacaccccgt tcacaaccac cacatagaag gtgaattagg cccagtgaac
     4921 tcccaccccc tgagaaccca caagctcaac ggaaaattcc caataggcaa gacctaccac
     4981 aagcttaaaa atcttaaggc cctctaaaag taaaagttgg cctcaccata caaagggacc
     5041 accctcccag agcagttcga ctggaggaat gtgaacggtt catcttacgt tccggaacct
     5101 tttgaccaaa gatcctgtgg gtcttgctac gcatgcgcct caatctatat gtacatgtca
     5161 agaattcgcg tgtctagcaa gaattagtag aaaccgaatc tttcaatcca gcacgctgtg
     5221 gattgcaatt tttactcgca gggctgtgcc ggcggattcg accagcaggt ctcgaagttt
     5281 gctgaggagt tcggtatcaa gcacgaaagc gactacaaac catattctgc agaagacgag
     5341 aagtgcgaag ccggcgaatt tttgtccggg aatcgtgagt ttttccaggc ctaccagccg
     5401 cttggtggct attacggcgc agtttatgac cccacggaaa tgcagtggga gctgcaccgc
     5461 aatggcccct ttgtggtttc cgttatggtg gaggggaact tccccgacga cccctacggc
     5521 gactaagaac caagcaaata cgctgaggag gaagacggtg aacgccatta ctacattagc
     5581 gaagcaaacc acgctgtttt gttgattggc tggaccacgc tgcatgacgg caatcctgcg
     5641 tggattattc agaactcctg ggggaaaagc tggaacggcg atggaacttt cttgctagcg
     5701 agagggtcta acgcatatgg tgttgaaagc taccctggca tcgcattctg gaggcccgat
     5761 ggtgctgtcc attaccctga ggttgttact gagactgcgc ttacctttgt aactgttgtt
     5821 ttgggagcac ttctttctgt ggtcactgct gttatgctcg ttttggcaat tatgctctgc
     5881 aaaaagaaca acgccagcgg gtataagaag attgaggacg ctgagcggcc gccctggtcc
     5941 cacccccaat tcgagaaggg agggggcagt ggtggcgggt ctggcggtgg atcgtggtca
     6001 catcctcagt ttgagaaggg agccagcggc gaagattaca aagatgacga cgacaagggg
     6061 ccctgattaa atttgatttt tttaaataaa ctattattcc aaaattttga tttatattat
     6121 ttggagctcc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     6181 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     6241 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     6301 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6361 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     6421 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6481 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6541 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6601 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6661 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6721 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     6781 g
//
