LOCUS       40924_13500        4908 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-Rluc-PHLEO, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4908)
  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  JOURNAL   Unpublished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 4908)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 4908)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4908)
  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            "Unpublished"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..4908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..13
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     CDS             14..946
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             998..1369
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    1373..1385
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(1467..1485)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1503..1519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1527..1543)
                     /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(1551..1581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1596..1617)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1905..2493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2634..3005)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(3215..4006)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4358..4657)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4795..4811
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4818..4836
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcctgcgtg gccatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac
       61 tgggcctcag tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta
      121 ctatgattcc gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc
      181 cagctacctg tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc
      241 tgatctgatc ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga
      301 tcactacaag tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt
      361 tgtgggccac gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa
      421 gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg
      481 gcctgacatc gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct
      541 tgagaataac ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc
      601 tgaggagttc gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac
      661 cctctcctgg cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat
      721 tgtccgcaac tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga
      781 gtccgaccct gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac
      841 cgagttcgtg aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg
      901 taagtacatc aagagcttcg tggagcgcgt gctgaagaac gagcagaccg gtggtgggag
      961 cggaggtggc ggatcaggtg gcggaggctc cggagggatg gccaagttga ccagtgccgt
     1021 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     1081 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     1141 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     1201 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     1261 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     1321 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gaggcctctg
     1381 tggcctcaag ggcgaattcc agcacactgg cggccgttac tagtggatcc gagctcggta
     1441 ccaagcttga tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca
     1501 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1861 cagcaaaagc ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2341 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttagcacgtg tcagtcctgc
     2641 tcctcggcca cgaagtgcac gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc
     2701 cacggctgct cgccgatctc ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac
     2761 acgacctccg accactcggc gtacagctcg tccaggccgc gcacccacac ccaggccagg
     2821 gtgttgtccg gcaccacctg gtcctggacc gcgctgatga acagggtcac gtcgtcccgg
     2881 accacaccgg cgaagtcgtc ctccacgaag tcccgggaga acccgagccg gtcggtccag
     2941 aactcgaccg ctccggcgac gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttc
     3001 gccatggtgg ccctcctcac gtgctattat tgaagcattt atcagggtta ttgtctcatg
     3061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3121 ccccgaaaag tgccacctgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3181 accgcatcag gaaattgtaa gcgttaataa ttcagaagaa ctcgtcaaga aggcgataga
     3241 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     3301 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     3361 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     3421 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg
     3481 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     3541 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     3601 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     3661 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     3721 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     3781 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg
     3841 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     3901 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     3961 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     4021 atcctgtctc ttgatcagag cttgatcccc tgcgccatca gatccttggc ggcgagaaag
     4081 ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt
     4141 ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc
     4201 aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     4261 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgaaa aggatctagg
     4321 tgaagatcct ttttgataat ctcatgcctg acatttatat tccccagaac atcaggttaa
     4381 tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg
     4441 agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca
     4501 ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg gccaggggga
     4561 tcaccatccg tcgccccggc gtgtcaataa tatcactctg tacatccaca aacagacgat
     4621 aacggctctc tcttttatag gtgtaaacct taaactgccg tacgtatagg ctgcgcaact
     4681 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     4741 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     4801 cgacggccag tgaattgtaa tacgactcac tatagggcga attgggccct ctagatgcat
     4861 gctcgagcgg ccgccagtgt gatggatatc tgcagaattc gcccttga
//
