LOCUS       40924_42724        7827 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Protein fusion vector pTcCFPH-NH 1.1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  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 7827)
  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 7827)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7827)
  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..7827
                     /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=CFP
                     /note="cyan variant of 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             4305..5327
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     promoter        complement(5639..5657)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5678..5694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5702..5718)
                     /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(5726..5756)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5771..5792)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6080..6668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6842..7699)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7700..7804)
                     /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 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     1621 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     1681 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     1741 caacacttgt cactactttg acttggggtg ttcaatgctt ttcaagatac ccagatcata
     1801 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     1861 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     1921 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     1981 gacacaaatt ggaatacaac tatatttcac acaatgtata catcactgca gacaaacaaa
     2041 agaatggaat caaagctcat ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     2101 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     2161 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     2221 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     2281 aatctagaat 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 cggtatgaaa aagcctgaac
     4321 tcaccgcgac gtctgtcgag aagtttctga tcggaaagtt cgacagcgtc tccgacctga
     4381 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     4441 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     4501 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga
     4561 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     4621 ccgaactgcc cgctgttctt cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     4681 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     4741 catggcgtga tttcatttgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     4801 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     4861 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     4921 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     4981 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     5041 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     5101 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     5161 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     5221 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     5281 ccgatagtgg aaaccgacgc ctcagcactc gtccgagggc aaaggaatag ctcgagtctg
     5341 cgatgctgtg gaccgacgca ttgaaataca caccgtcttt ggcgttcctt tttttttttt
     5401 tgatttgttt tttttattga gatgttttgt tttgtttttt tcagttttta ttatatcagc
     5461 agtttgtccg cctgcattca tgcagtgttt ggtaattctt tctatttttt ggaattatgg
     5521 cgataatatt cttgtcttta aacttcttat aaccaattgt gctttagagt ttcctgctta
     5581 gttgctatta acacactgtt aggaacgtga aaccgggccc ggtacccagc ttttgttccc
     5641 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5701 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5761 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5821 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5881 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5941 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6001 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6481 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7801 cgcgcacatt tccccgaaaa gtgccac
//
