LOCUS       40924_41141        4296 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Episomal vector pSpiro-PAC-alphaTub-3xHA-N, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4296)
  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 4296)
  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 4296)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4296)
  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..4296
                     /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..2423)
                     /note="ornithine carbamoyltransferase from Spironucleus 
                     salmonicida"
     CDS             complement(2430..3023)
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     5'UTR           complement(3030..3191)
                     /note="ornithine carbamoyltransferase from Spironucleus 
                     salmonicida"
     5'UTR           3204..3446
                     /label=alpha tubulin from Spironucleus salmonicida
                     /note="alpha tubulin from Spironucleus salmonicida"
     CDS             3453..3533
                     /label=3xHA
                     /note="three tandem HA epitope tags"
     3'UTR           3579..3638
                     /note="ribosomal protein S15A from Spironucleus
                     salmonicida"
     promoter        complement(3653..3671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3681..3697)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3839..4294
                     /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 agtgttgtat
     2221 ggaatattta tatataaatg cattattaat aatgatatag tatcttaata tacattaata
     2281 atttgacaat tccaaacaaa aaagtaacat tattacaagt aataaagtat atattcatta
     2341 caaatatatt tcagataagt agtaactgtc tattcattat acaaatgagt aataaaagat
     2401 aattaatgaa atttagctga tcactcgagg gcaccgggct tgcgggtcat gcaccaggtc
     2461 gcgcggtcct tcgggcactc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag
     2521 gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg ctcggccgcc
     2581 tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg
     2641 ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc
     2701 atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg
     2761 gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg
     2821 tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc
     2881 agctcggtga cccgctcgat gtggcggtcc gggtcgacgg tgtggcgcgt ggcggggtag
     2941 tcggcgaacg cggcggcgag ggtgcgtacg gcccggggga cgtcgtcgcg ggtggcgagg
     3001 cgcaccgtgg gcttgtactc ggtgcccatg gtgggaagtg gtgaccaaat ctagaaaaaa
     3061 gatttaagaa tattatttgt taaattatta atctctatta tttccccctc cattatgttc
     3121 aaaaatatat ataaatatac cataattatt ctacttattt tgtgattttc ttaaatatgt
     3181 gagtatgcct ccatatgaag cttataatta tagctctagt atagagttac actaaaaatt
     3241 acgatttagc atattaatat taaaaaaaca caatttttat atggaaaaat gataaatttt
     3301 tataaaagtt ttatatgcaa caatatatat cttcagttta ccagaaattc tattgaattt
     3361 attatatgca tgtctaaatt tagtctaaaa caaacgcgca aactataaaa cttgcaaaat
     3421 ttcctttcct tcaaattacc atgagggata tctatcccta cgacgtgcca gattacgcat
     3481 atccgtacga tgttcctgac tatgcttacc cctatgatgt ccctgactac gccggatcct
     3541 gcagtctaga cgcgtcgaat tcgcggccgc ccgggccctg attaaatttg atttttttaa
     3601 ataaactatt attccaaaat tttgatttat attatttgga gctccaattc gccctatagt
     3661 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3721 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3781 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     3841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3901 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3961 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4021 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4141 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4201 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4261 gcgaatttta acaaaatatt aacgcttaca atttag
//
