LOCUS       40924_13475        5622 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-luc2-PHLEO, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     A dual luciferase system for analysis of post-transcriptional gene 
            expression in Leishmania
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5622)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5622)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5622)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-MAY-2013) Biochemistry "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..13
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     CDS             14..1663
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             1712..2083
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    2087..2099
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(2181..2199)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2217..2233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2241..2257)
                     /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(2265..2295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2310..2331)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2619..3207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3348..3719)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(3929..4720)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5072..5371)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5509..5525
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5532..5550
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcctgcgtg gccatggaag atgccaaaaa cattaagaag ggcccagcgc cattctaccc
       61 actcgaagac gggaccgccg gcgagcagct gcacaaagcc atgaagcgct acgccctggt
      121 gcccggcacc atcgccttta ccgacgcaca tatcgaggtg gacattacct acgccgagta
      181 cttcgagatg agcgttcggc tggcagaagc tatgaagcgc tatgggctga atacaaacca
      241 tcggatcgtg gtgtgcagcg agaatagctt gcagttcttc atgcccgtgt tgggtgccct
      301 gttcatcggt gtggctgtgg ccccagctaa cgacatctac aacgagcgcg agctgctgaa
      361 cagcatgggc atcagccagc ccaccgtcgt attcgtgagc aagaaagggc tgcaaaagat
      421 cctcaacgtg caaaagaagc taccgatcat acaaaagatc atcatcatgg atagcaagac
      481 cgactaccag ggcttccaaa gcatgtacac cttcgtgact tcccatttgc cacccggctt
      541 caacgagtac gacttcgtgc ccgagagctt cgaccgggac aaaaccatcg ccctgatcat
      601 gaacagtagt ggcagtaccg gattgcccaa gggcgtagcc ctaccgcacc gcaccgcttg
      661 tgtccgattc agtcatgccc gcgaccccat cttcggcaac cagatcatcc ccgacaccgc
      721 tatcctcagc gtggtgccat ttcaccacgg cttcggcatg ttcaccacgc tgggctactt
      781 gatctgcggc tttcgggtcg tgctcatgta ccgcttcgag gaggagctat tcttgcgcag
      841 cttgcaagac tataagattc aatctgccct gctggtgccc acactattta gcttcttcgc
      901 taagagcact ctcatcgaca agtacgacct aagcaacttg cacgagatcg ccagcggcgg
      961 ggcgccgctc agcaaggagg taggtgaggc cgtggccaaa cgcttccacc taccaggcat
     1021 ccgccagggc tacggcctga cagaaacaac cagcgccatt ctgatcaccc ccgaagggga
     1081 cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc gaggctaagg tggtggactt
     1141 ggacaccggt aagacactgg gtgtgaacca gcgcggcgag ctgtgcgtcc gtggccccat
     1201 gatcatgagc ggctacgtta acaaccccga ggctacaaac gctctcatcg acaaggacgg
     1261 ctggctgcac agcggcgaca tcgcctactg ggacgaggac gagcacttct tcatcgtgga
     1321 ccggctgaag agcctgatca aatacaaggg ctaccaggta gccccagccg aactggagag
     1381 catcctgctg caacacccca acatcttcga cgccggggtc gccggcctgc ccgacgacga
     1441 tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca tgaccgagaa
     1501 ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc aagaagctgc gcggtggtgt
     1561 tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag ttggacgccc gcaagatccg
     1621 cgagattctc attaaggcca agaagggcgg caagatcgcc gtgggtggtg ggagcggagg
     1681 tggcggatca ggtggcggag gctccggagg gatggccaag ttgaccagtg ccgttccggt
     1741 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     1801 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     1861 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     1921 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     1981 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     2041 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgaggcc tctgtggcct
     2101 caagggcgaa ttccagcaca ctggcggccg ttactagtgg atccgagctc ggtaccaagc
     2161 ttgatgcata gcttgagtat tctatagtgt cacctaaata gcttggcgta atcatggtca
     2221 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2281 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2581 aagcccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3301 ttcacctaga tccttttaaa ttaaaaatga agttttagca cgtgtcagtc ctgctcctcg
     3361 gccacgaagt gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg cccccacggc
     3421 tgctcgccga tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc
     3481 tccgaccact cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg
     3541 tccggcacca cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca
     3601 ccggcgaagt cgtcctccac gaagtcccgg gagaacccga gccggtcggt ccagaactcg
     3661 accgctccgg cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa cttcgccatg
     3721 gtggccctcc tcacgtgcta ttattgaagc atttatcagg gttattgtct catgagcgga
     3781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3841 aaagtgccac ctgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3901 tcaggaaatt gtaagcgtta ataattcaga agaactcgtc aagaaggcga tagaaggcga
     3961 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     4021 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     4081 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     4141 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     4201 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     4261 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     4321 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     4381 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     4441 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     4501 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     4561 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     4621 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     4681 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     4741 tctcttgatc agagcttgat cccctgcgcc atcagatcct tggcggcgag aaagccatcc
     4801 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     4861 cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta
     4921 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     4981 tccggggtca gcaccgtttc tgcggactgg ctttctacgt gaaaaggatc taggtgaaga
     5041 tcctttttga taatctcatg cctgacattt atattcccca gaacatcagg ttaatggcgt
     5101 ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata acggagaccg
     5161 gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata tgcaccaccg
     5221 ggtaaagttc acgggagact ttatctgaca gcagacgtgc actggccagg gggatcacca
     5281 tccgtcgccc cggcgtgtca ataatatcac tctgtacatc cacaaacaga cgataacggc
     5341 tctctctttt ataggtgtaa accttaaact gccgtacgta taggctgcgc aactgttggg
     5401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     5461 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     5521 ccagtgaatt gtaatacgac tcactatagg gcgaattggg ccctctagat gcatgctcga
     5581 gcggccgcca gtgtgatgga tatctgcaga attcgccctt ga
//
