LOCUS       40924_25736        6737 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pIS-HGFPna, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  AUTHORS   Morelle C, Sterkers Y, Crobu L, MBang-Benet DE, Kuk N, Portales P, 
            Bastien P, Pages M, Lachaud L.
  TITLE     The nucleoporin Mlp2 is involved in chromosomal distribution during 
            mitosis in trypanosomatids
  JOURNAL   Nucleic Acids Res. 43 (8), 4013-4027 (2015)
  PUBMED    25690889
REFERENCE   2  (bases 1 to 6737)
  AUTHORS   Crobu L, Sterkers Y, Bastien P, Pages M.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-FEB-2014) Departement of Parasitology-Mycology UMR 
            MIVEGEC (CNRS 5290 - IRD 224 - University Montpellier 1 and 2), 
            C.H.R.U. de Montpellier, 39 Avenue Charles Flahault, Montpellier 
            Cedex 5, Languedoc-Roussillon F34295, France
REFERENCE   3  (bases 1 to 6737)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6737)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2015"; volume: "43"; issue: "8"; pages: 
            "4013-4027"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-FEB-2014) Departement of Parasitology-Mycology UMR MIVEGEC (CNRS
            5290 - IRD 224 - University Montpellier 1 and 2), C.H.R.U. de 
            Montpellier, 39 Avenue Charles Flahault, Montpellier Cedex 5, 
            Languedoc-Roussillon F34295, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /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"
     CDS             1272..2294
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2643..3359
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3375..3392
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(3541..3552)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     promoter        complement(4552..4570)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4591..4607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4615..4631)
                     /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(4639..4669)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4684..4705)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4993..5581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5755..6612)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6613..6717)
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgc
      661 atcgacatcg cgcacgtgtt tgcggtgtac tgtgcgtcga tgatcgagcg gccaagcgtc
      721 cagcttcacg cgaggaggac tgcatcgctg ctcctgtgcg cgtgcgtgca cgcgtcaagg
      781 cacgctcgat tcgttcccag ggtggatacc ctcccccctc ccccagctgc agacaacccc
      841 caccctctcg aggcttgcac ccaggctcgt cagtctctct accactttct gccttctgta
      901 gtgcacccca cccctgcatt ctcctctctc tcacacacac acacacacac acacacacac
      961 acacacacac acgtacacac gtacacacac gtgcgcatgc ttgcacgcat acgcgacgaa
     1021 ttcgaaagct cacctcattc ctccctcctc acaccatcat cggcatccat agagacacgt
     1081 gcgcgtagaa cgatacagct cacgcgcaca gagaggcgtg ttctggtcgt gcgcatcatc
     1141 ggcacagcac tggcggacga aactcgcaca caggcacgcc gcctcctttc acccgtcata
     1201 gatagttgaa ttagacgccc tcctcctccc tcatcatcgc cgtcgtcatc cgggtccgag
     1261 cactagtatg gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     1321 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     1381 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     1441 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     1501 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     1561 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     1621 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     1681 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     1741 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     1801 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     1861 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     1921 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     1981 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     2041 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     2101 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     2161 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     2221 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     2281 cgagggcaaa ggaataggga tccaaagtgt gacaacgtcg caccatgtgt aggttttcat
     2341 ttatgttctt tctttctttc ttttgtgaat ttgttttctg tctcaaatgt ttttaattcg
     2401 cttgggacct atgtttttct tgtttttttg ctcacccttt gtgtaggagg caccctgtca
     2461 cgtctgtggt tgcgtgtatg ccttccttcc ccttattcgc ttcttcctgt cgtgtcacac
     2521 ctcnttctcc ctctcccttt ccgccttttc tttcaatctt gttttctcga ccagccctac
     2581 tagaggagaa agaatagtaa ccctttcatc aaagaaaata gttcaaacga attggtttaa
     2641 acatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2701 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2761 acggcaagct taccctcaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2821 ccctcgtgac caccttcacc tacggcgtgc agtgctttag ccgctacccc gaccatatga
     2881 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2941 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3001 tggtgaaccg catcgagctc aagggcatcg atttcaagga ggacggcaac atcctggggc
     3061 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3121 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3181 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3241 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3301 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc
     3361 aattgcccgt taaccaccat caccatcatc actagagtag atgccgaccg ggatcgatcc
     3421 tctagtagtg atcccgcccg cacccgcgcg cgctcaggcc gcgtgtcgtc gtgctctttc
     3481 catttttttt ttgtgtgtgg tggtgatttg agctgctctc cgttgtgtgc tggggaccct
     3541 ccttccctcg atctcctcgt gcggggtctc cgacgccgca gacgcggtgc gtgcgagcgt
     3601 gcacgtggct gtcgccgtgg ctgtagttgc gagcggagag agagagagag ggagggaggg
     3661 agggcagagg gcagaggaca tgcgggtggg aacgtgcacc ggcctcgtct cacgcagctg
     3721 gagcccacga atgccaccac cacacccttc ttcccccccc ctccctcttc ttcccacggc
     3781 ggcgacgacg acggggctca gctcacgctt ctgtagggta ttatattaaa gcacatgtgc
     3841 gtgctgtgct tcggcttctt gttggtggtc gttcgctttc aagtccacga ctcctgccgt
     3901 tgctcaccgc cgggctgtct tctttcgccc ctccctcggt gcctcttccg ccgccgcacg
     3961 cgtgcgctga tcacgcgtct tgcgtgatgt gcctgtgtgt atacacaccg tgcacggaga
     4021 gagcgagcga gcgagagaga gagagggcga gaggaagaat ggccggtgcc tcgcgtgggc
     4081 aagcgtgcgc gagtttgtcg tgtcgctgcg ccgcgtggat gtcgcgcagc gcaagacccc
     4141 gtgcgcctga aacgtgaagg gaggtcgaga agcgtgccgc atgaggcctt aaggcaggaa
     4201 agtgaaaaga gctcgacgcg agtgcagcac gcgcgcatcg acaggacgga atagacactc
     4261 agccccttcc cctttcaaaa actgaatgga cggacatcgt aacgcgcttc tctcccctcc
     4321 actccccctg tccctgtctc cttgacgtgt gctgctctcg tgagcgacag atgaagcgag
     4381 gcgtagaagg aaaggggggg ggagcaggcc tgcacacaca cacacacaca cacacacaca
     4441 cacacacaca cacacacaca cacacgaaaa gcaagtttga ggcagacgtg gtttcacacg
     4501 attcctgtgt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     4561 agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4621 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4681 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4741 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4801 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4861 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4921 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4981 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5041 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5101 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5161 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5221 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5281 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5341 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5401 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5461 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5521 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5581 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5641 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5701 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5761 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5821 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5881 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5941 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6001 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6061 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6121 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6181 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6241 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6301 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6361 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6421 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6481 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6541 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6601 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6661 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6721 atttccccga aaagtgc
//
