LOCUS       40924_42754        7596 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Protein fusion vector pTcGFPN-NH 1.1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7596)
  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 7596)
  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 7596)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7596)
  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..7596
                     /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             1569..2282
                     /label=EGFP
                     /note="enhanced GFP"
     protein_bind    2292..2411
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2441..2471
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2525..3181
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             3526..3828
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3872..3996)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4308..5096
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(5408..5426)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5447..5463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5471..5487)
                     /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(5495..5525)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5540..5561)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5849..6437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6611..7468)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7469..7573)
                     /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 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1621 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1681 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1741 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1801 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1861 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1921 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1981 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2041 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2101 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2161 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2221 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2281 agtctagaat cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata
     2341 tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac
     2401 atatccagtc atattggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg
     2461 ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc
     2521 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     2581 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     2641 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     2701 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     2761 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     2821 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     2881 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     2941 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     3001 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     3061 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     3121 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     3181 gtaaacgcgt ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga
     3241 tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat
     3301 gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca
     3361 tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc
     3421 tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta
     3481 ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt
     3541 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
     3601 gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa
     3661 gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc
     3721 accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac
     3781 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
     3841 tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca
     3901 gcattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
     3961 ttacgtttct cgttcagctt tcttgtacaa agtggtgata ctagtccggc gcgccattga
     4021 tctgcgatgc tgtggaccga cgcattgaaa tacacaccgt ctttggcgtt cctttttttt
     4081 tttttgattt gtttttttta ttgagatgtt ttgttttgtt tttttcagtt tttattatat
     4141 cagcagtttg tccgcctgca ttcatgcagt gtttggtaat tctttctatt ttttggaatt
     4201 atggcgataa tattcttgtc tttaaacttc ttataaccaa ttgtgcttta gagtttcctg
     4261 cttagttgct attaacacac tgttaggaac gtgaaaccac cggtatgatt gaacaagatg
     4321 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4381 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4441 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4501 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4561 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4621 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4681 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4741 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4801 cgccagccga actgttcgcc aggctcaagg cgcgaatgcc cgacggcgat gatctcgtcg
     4861 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4921 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4981 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5041 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgac
     5101 tcgagtctgc gatgctgtgg accgacgcat tgaaatacac accgtctttg gcgttccttt
     5161 tttttttttt gatttgtttt ttttattgag atgttttgtt ttgttttttt cagtttttat
     5221 tatatcagca gtttgtccgc ctgcattcat gcagtgtttg gtaattcttt ctattttttg
     5281 gaattatggc gataatattc ttgtctttaa acttcttata accaattgtg ctttagagtt
     5341 tcctgcttag ttgctattaa cacactgtta ggaacgtgaa accgggcccg gtacccagct
     5401 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     5461 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5521 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5581 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5641 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5701 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5761 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5821 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5881 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5941 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6001 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6061 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6121 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6181 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6241 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6301 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6361 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6421 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6481 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6541 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6601 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6661 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6721 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6781 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6841 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6901 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6961 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7021 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7081 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7141 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7201 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7261 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7321 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7381 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7441 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7501 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7561 taggggttcc gcgcacattt ccccgaaaag tgccac
//
