LOCUS       40924_42749        7589 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Protein fusion vector pTcGFPN-CO 1.1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  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 7589)
  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 7589)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7589)
  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..7589
                     /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"
     protein_bind    1569..1688
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1718..1748
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1802..2458
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2803..3105
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3149..3273)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3281..3997
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             4301..5089
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(5401..5419)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5440..5456)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5464..5480)
                     /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(5488..5518)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5533..5554)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5842..6430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6604..7461)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7462..7566)
                     /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 cgtgaaacct
     1561 ctagaatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
     1621 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     1681 tccagtcata ttggcggccg cattaggcac cccaggcttt acactttatg cttccggctc
     1741 gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa
     1801 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1861 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1921 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1981 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     2041 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2101 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2161 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2221 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2281 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2341 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2401 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2461 aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     2521 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct
     2581 atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat
     2641 atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc
     2701 gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg
     2761 aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac
     2821 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     2881 acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     2941 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     3001 gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc
     3061 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     3121 cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagca
     3181 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     3241 cgtttctcgt tcagctttct tgtacaaagt ggtgatacta gtaatgagca agggcgagga
     3301 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3361 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3421 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3481 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3541 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3601 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3661 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3721 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     3781 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3841 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     3901 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3961 cgccgggatc actctcggca tggacgagct gtacaagtga ggcgcgccat tgatctgcga
     4021 tgctgtggac cgacgcattg aaatacacac cgtctttggc gttccttttt ttttttttga
     4081 tttgtttttt ttattgagat gttttgtttt gtttttttca gtttttatta tatcagcagt
     4141 ttgtccgcct gcattcatgc agtgtttggt aattctttct attttttgga attatggcga
     4201 taatattctt gtctttaaac ttcttataac caattgtgct ttagagtttc ctgcttagtt
     4261 gctattaaca cactgttagg aacgtgaaac caccggtatg attgaacaag atggattgca
     4321 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4381 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4441 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4501 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4561 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4621 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4681 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4741 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4801 cgaactgttc gccaggctca aggcgcgaat gcccgacggc gatgatctcg tcgtgaccca
     4861 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4921 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4981 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5041 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gactcgagtc
     5101 tgcgatgctg tggaccgacg cattgaaata cacaccgtct ttggcgttcc tttttttttt
     5161 tttgatttgt tttttttatt gagatgtttt gttttgtttt tttcagtttt tattatatca
     5221 gcagtttgtc cgcctgcatt catgcagtgt ttggtaattc tttctatttt ttggaattat
     5281 ggcgataata ttcttgtctt taaacttctt ataaccaatt gtgctttaga gtttcctgct
     5341 tagttgctat taacacactg ttaggaacgt gaaaccgggc ccggtaccca gcttttgttc
     5401 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5461 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5521 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5581 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5641 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7561 tccgcgcaca tttccccgaa aagtgccac
//
