LOCUS       40924_42814        7359 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Protein fusion vector pTcPTPN-NH 1.1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  AUTHORS   Kugeratski FG, Batista M, Inoue AH, Ramos BD, Krieger MA, Marchini/ 
            FK.
  TITLE     pTcGW plasmid vectors 1.1 version: a versatile tool for Trypanosoma 
            cruzi gene characterisation
  JOURNAL   Mem. Inst. Oswaldo Cruz (2015) In press
  PUBMED    26200713
REFERENCE   2  (bases 1 to 7359)
  AUTHORS   Kugeratski FG, Batista M, Inoue AH, Ramos BD, Krieger MA, Marchini 
            FK.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2015) Mass Spectrometry Platform, Instituto Carlos
            Chagas, Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 
            81350010, Brasil
REFERENCE   3  (bases 1 to 7359)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7359)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mem. Inst. 
            Oswaldo Cruz (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-APR-2015) Mass Spectrometry Platform, Instituto Carlos Chagas, 
            Professor Algacyr Munhoz Mader, 3775, Curitiba, Parana 81350010, 
            Brasil"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..1275
                     /note="derived from Trypanosoma cruzi 18S ribosomal RNA
                     gene promoter region"
                     /regulatory_class="promoter"
     CDS             1599..1772
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
     CDS             1776..1946
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             1983..2003
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     protein_bind    2055..2174
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2204..2234
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2288..2944
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             3289..3591
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3635..3759)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4071..4859
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(5171..5189)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5210..5226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5234..5250)
                     /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(5258..5288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5303..5324)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5612..6200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6374..7231)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7232..7336)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccagtttcct tgcggggctt atttggtgtg ttggattttt aattttttga tgattcggag
      721 gaattttttg caatttttgg gtacccctaa aaaaattttt ggagtttccg gggttcgatc
      781 caacttgtgc ttttttaacc gtccatttgt gtctagtaca tcatggagcg gtattctctc
      841 cggtggccca ggtggcgtcg gaaggaattg gtgtccctca ggatgtttgg tttttcgttt
      901 ttttttttgc ggtcttttgt ttcgcatcgc cgtatcgtgt gcggcgcgac cttctttttg
      961 gctcattggt cagtgaggtt tataataaag tggatcattt gggtttttgg ccctttgcgg
     1021 cgcacccgcg gaatggggga gggattttca tataatgtag tgtacattgt gtgtgggtcg
     1081 caaaatgtca tgccgtgtgc aatgtttatt tgcgtgctgt gcgtgtcctt ttgtgattcc
     1141 accacttgtg tgtattttac cttatcgtct ctccctgccg ttggctgtgc gtgaattgtg
     1201 cttcgcattc gtgggaaatc aaaaggggaa tcaacgtgtt cttttttttc agaaggggag
     1261 tcaaagcgcc gtgtggatat ctctgcgatg ctgtggaccg acgcattgaa atacacaccg
     1321 tctttggcgt tccttttttt ttttttgatt tgtttttttt attgagatgt tttgttttgt
     1381 ttttttcagt ttttattata tcagcagttt gtccgcctgc attcatgcag tgtttggtaa
     1441 ttctttctat tttttggaat tatggcgata atattcttgt ctttaaactt cttataacca
     1501 attgtgcttt agagtttcct gcttagttgc tattaacaca ctgttaggaa cgtgaaacca
     1561 tggcatgcat ggcaggcctt gcgcaccacg atgaagccgt ggacaacaaa ttcaacaaag
     1621 aacaacaaaa cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa
     1681 acgccttcat ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag
     1741 ctaaaaagct aaatgatgct caggcgccga aagtagacaa caaattcaac aaagaacaac
     1801 aaaacgcgtt ctatgagatc ttacatttac ctaacttaaa cgaagaacaa cgaaacgcct
     1861 tcatccaaag tttaaaagat gacccaagcc aaagcgctaa ccttttagca gaagctaaaa
     1921 agctaaatga tgctcaggcg ccgaaagtag acgcgaattg tgatatacct acaactgctt
     1981 ctgaaaattt atattttcaa ggtgaactag aagatcaggt ggatcctcgt cttattgatg
     2041 ggaaatctag aatcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
     2101 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     2161 aacatatcca gtcatattgg cggccgcatt aggcacccca ggctttacac tttatgcttc
     2221 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
     2281 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     2341 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     2401 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     2461 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     2521 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     2581 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     2641 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     2701 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     2761 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     2821 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     2881 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     2941 ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     3001 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     3061 tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     3121 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     3181 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     3241 ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag
     3301 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     3361 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     3421 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     3481 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     3541 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     3601 ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     3661 acagcattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     3721 attttacgtt tctcgttcag ctttcttgta caaagtggtg atactagtcc ggcgcgccat
     3781 tgatctgcga tgctgtggac cgacgcattg aaatacacac cgtctttggc gttccttttt
     3841 ttttttttga tttgtttttt ttattgagat gttttgtttt gtttttttca gtttttatta
     3901 tatcagcagt ttgtccgcct gcattcatgc agtgtttggt aattctttct attttttgga
     3961 attatggcga taatattctt gtctttaaac ttcttataac caattgtgct ttagagtttc
     4021 ctgcttagtt gctattaaca cactgttagg aacgtgaaac caccggtatg attgaacaag
     4081 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4141 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4201 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4261 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4321 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4381 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4441 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4501 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4561 tcgcgccagc cgaactgttc gccaggctca aggcgcgaat gcccgacggc gatgatctcg
     4621 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4681 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4741 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4801 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4861 gactcgagtc tgcgatgctg tggaccgacg cattgaaata cacaccgtct ttggcgttcc
     4921 tttttttttt tttgatttgt tttttttatt gagatgtttt gttttgtttt tttcagtttt
     4981 tattatatca gcagtttgtc cgcctgcatt catgcagtgt ttggtaattc tttctatttt
     5041 ttggaattat ggcgataata ttcttgtctt taaacttctt ataaccaatt gtgctttaga
     5101 gtttcctgct tagttgctat taacacactg ttaggaacgt gaaaccgggc ccggtaccca
     5161 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     5221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7321 aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
