LOCUS       40924_44404        6355 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pTtag, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6355)
  AUTHORS   El-Haddad H, Przyborski JM, Kraft LG, McFadden GI, Waller RF, Gould 
            SB.
  TITLE     Characterization of TtALV2, an Essential Charged Repeat Motif 
            Protein of the Tetrahymena thermophila Membrane Skeleton
  JOURNAL   Eukaryotic Cell 12 (6), 932-940 (2013)
  PUBMED    23606287
REFERENCE   2  (bases 1 to 6355)
  AUTHORS   Gould SB, McFadden GI, Waller RF.
  TITLE     Analysis of the two alveolate-characterizing epiplasmin proteins 
            alveolin 1 and 2 from Tetrahymena thermophila
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6355)
  AUTHORS   Gould SB, McFadden GI, Waller RF.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-FEB-2009) School of Botany, University of Melbourne, 
            Professors Walk, Parkville, VIC 3010, Australia
REFERENCE   4  (bases 1 to 6355)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6355)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Eukaryotic 
            Cell"; date: "2013"; volume: "12"; issue: "6"; pages: "932-940"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (24-FEB-2009) School of Botany, University of Melbourne, Professors 
            Walk, Parkville, VIC 3010, Australia"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           6..520
                     /label=BTU1
                     /note="BTU1"
     regulatory      527..1455
                     /label=MTT promoter
                     /note="MTT promoter"
                     /regulatory_class="promoter"
     misc_feature    1456..1494
                     /label=MCS
                     /note="MCS"
     CDS             2221..2247
                     /label=9xHis
                     /note="9xHis affinity tag"
     3'UTR           2257..3490
                     /label=BTU1
                     /note="BTU1"
     promoter        complement(3509..3527)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3548..3564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3572..3588)
                     /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(3596..3626)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3641..3662)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3950..4538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4712..5569)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5570..5674)
                     /label=AmpR promoter
     rep_origin      complement(5700..6155)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6297..6313
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6323..6341
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcccgatc tctctctttc tatcgtattt tgcaataata ggtattaact tttatactga
       61 ttgttagtag atgccttcaa attttctttt tatttaaatt catatgctat atcttttaaa
      121 acactccaca ttttattgtt gctaactgtg ctattgatct ttaagtcaat agctgctcat
      181 tttgttgaac tccacagaga cactaaattt gtttattttg atggatgctt tataattaaa
      241 gttacgtaat ctgcttgaca tttagccaac tatataaaaa agatcaaaat gtagcttaaa
      301 tctcaaaaaa tcatcataat ttactatcaa attattaaga aattcatata atcaccactt
      361 tattgacttt tattcatctt atagagtgat agtagagttg agccaaattg atacttgttt
      421 acgttgtatt attttgaaat tttaaaaaaa tgaaaatgag agaaaaattt atttaaattt
      481 gagcttagaa tctttaagga agatcaaaaa tgggctaact agatcttaag tcagtagaat
      541 tttagatatt ttcatagatt ttttaataag tagacaattt atttctaaaa aatatttaaa
      601 aataaaaaat aataagggtt ttgaataact cctttaattt aaatacacat ttttaaattt
      661 tttttagctc tttaaatatt cataaaaata aaaaataact aactaaaaat aaataaaaag
      721 ataataatga ttaaaggtat aatactgaat aagaaaaaac ataatagagt acttattttt
      781 tatatcacta tttttaatat cttgaaagca aaactttttt atatatctta aaatatattg
      841 tatcgtttat tcaattattt tctttaaatt tcaaatatat tgataaaaaa gatgacatgt
      901 tttttaaaga aaacatgaaa tataaaatag ataaatatca attattttat ttattaaata
      961 tataagctgc tcaaaacata gctcactcat caattataat atgtgaatca ttaattttca
     1021 aaatattact cattatttag gctatcattt attttttatt ttcaattatc cgtttctatt
     1081 atattttaat attaagttgt gattcttgag ttttgtgtca tgaattattt gtaaatcttt
     1141 ttatttctga taaaaaatat aaattgattg actcatgatt taaatcatga gtcaacctaa
     1201 ctaattttca aaattcttct attctaaaat atagatgtga ttcttgaatc tctcttgaat
     1261 ataaagtaat tttttatatt tctgatataa ttcttagcta cgtgattcac gatttatgca
     1321 atgatccata taaaataatg taaatagtgt atatatatat attcgtcttt tttattcttt
     1381 atataattta aaaaaattaa aaaaatttaa taaagctcta ataaaataaa taataatact
     1441 aaacttaaaa taatgaagct tgctgcatgc gctctcgagg ctacgcgttc tagaatggtt
     1501 tcaaagggtg aagaactttt cactggtgtt gttcctattc ttgttgaact tgatggtgat
     1561 gttaacggtc acaagttctc agtttccggt gaaggtgaag gtgatgctac ttacggtaag
     1621 ttaactctta agttcatttg tactactggt aagttacctg ttccttggcc tactcttgtt
     1681 actactctta cttatggtgt ttaatgcttc tcaagatacc ctgatcacat gaagtaacac
     1741 gatttcttca agtcagctat gcctgaaggt tatgtttaag aaagaactat tttcttcaag
     1801 gatgatggta attataagac tagagctgaa gttaagttcg aaggtgatac tcttgttaat
     1861 agaattgaac ttaagggtat tgatttcaag gaagatggta atattcttgg tcacaagtta
     1921 gaatataatt ataattcaca caatgtttat attatggctg ataagtaaaa gaatggtatt
     1981 aaggttaatt tcaagattag acacaatatt gaagatggtt cagtttaact tgctgatcac
     2041 tattaataaa atactcctat tggtgatggt cctgttcttc ttcctgataa tcactatctt
     2101 tcaacttaat cagctctttc aaaagatcct aatgaaaaga gagatcacat ggttcttctt
     2161 gaatttgtta ctgctgctgg tattactctt ggtatggatg aactttataa atctagaatg
     2221 catcatcacc atcaccacca tcaccacatg cattgaggat cctgagcgaa ctgaatcggt
     2281 cagctaaacc aaccaatcaa cataataaac tttattattt ttacttaagc atcttactgt
     2341 tgttgtaata gtagagaaag aaatacccaa ttaacttcat tcacataaca ttaatatcta
     2401 taaacatctt ttttctcaca tatatacaac tctctaaatc aacaaataac tttttaaaaa
     2461 taatggatat atattaacaa aataatatat ctctttttac aaaatagttc ttatataaat
     2521 acgtattctg cactcacccg catttttcac aacaaaaaca taccaaaaaa attcttactt
     2581 ctacatgttt cctttcttat tattacaaaa ttattttata aatagcatac aaaaataaat
     2641 acaataaaaa aataaacaaa atcctttttt attttgaatt atttaaaaca aatattttca
     2701 atcaatcagt cagtcagcat aatattaaag caacaaaaca aacccaagtt gtttttatag
     2761 ttttttaatt gcttttcagt actataaata aatttgttat tacttcaaga ttgataaact
     2821 tcttttttaa attaaatatc tatgaatgaa taaataagtt gatatctctt tttaacttgt
     2881 tttcctctct tttacttact tgccaatttt ttttttaaat taaagaaata tctttttatt
     2941 tttcaaaaac aaaatttatt ttcccttgta tacaaaaacc cctttattta aataaaatct
     3001 ttatgcccat caatagccac atctttttga attttattgc ttatactgcc tttcttttct
     3061 tttcttatga taatacttct tttttctatg attattggaa tatctttttt ttcttaatcg
     3121 tttttaaata aaattaagcg gcaacatatt ttattttctt aaaaatataa tattttataa
     3181 gtaataaaat taaaatttat attcaattgg tttctgattt tattttattc actattccaa
     3241 tagattgctt ttatttttaa aactagtttg tttatttatt taataaattt ttaatacttc
     3301 tttgaatttt tatatcttag ttctttaaaa agcgacttat agcaaaataa aatgtataac
     3361 taggaatagc tgaaaaaatc tttaatattt aaattattta aataggaaat atttaaaaag
     3421 taatgttttt gaatttttca aaattaaaca taaataatat gctttcttaa attttaattt
     3481 agactatttt gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     3541 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3601 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3661 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3721 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3781 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3841 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3901 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3961 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4021 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4081 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4141 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4201 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4261 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4321 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4381 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4441 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4501 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4561 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4621 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4681 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4741 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4801 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4861 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4921 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4981 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5041 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5101 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5161 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5221 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5281 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5341 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5401 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5461 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5521 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5581 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5641 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     5701 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5761 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5821 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5881 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5941 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6001 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6061 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6121 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     6181 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6241 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     6301 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtacc
//
