LOCUS       40924_13485        5559 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-Rluc-HYG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  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  JOURNAL   Unpublished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 5559)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5559)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5559)
  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            "Unpublished"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..5559
                     /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..2020
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    2024..2036
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(2118..2136)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2154..2170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2178..2194)
                     /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(2202..2232)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2247..2268)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2556..3144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3285..3656)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(3866..4657)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5009..5308)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5446..5462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5469..5487
                     /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 aaaaagcctg aactcaccgc
     1021 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     1081 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     1141 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     1201 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
     1261 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     1321 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     1381 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     1441 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     1501 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     1561 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     1621 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     1681 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     1741 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     1801 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     1861 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     1921 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     1981 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagggcctct gtggcctcaa
     2041 gggcgaattc cagcacactg gcggccgtta ctagtggatc cgagctcggt accaagcttg
     2101 atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc atggtcatag
     2161 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2221 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2281 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2341 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2401 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2461 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2521 cccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2581 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2641 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2701 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2761 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2821 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2881 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2941 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3001 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3061 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3121 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3241 acctagatcc ttttaaatta aaaatgaagt tttagcacgt gtcagtcctg ctcctcggcc
     3301 acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc
     3361 tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc
     3421 gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc
     3481 ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg
     3541 gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc
     3601 gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt cgccatggtg
     3661 gccctcctca cgtgctatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3721 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3781 gtgccacctg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3841 ggaaattgta agcgttaata attcagaaga actcgtcaag aaggcgatag aaggcgatgc
     3901 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     3961 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     4021 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4081 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4141 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     4201 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4261 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4321 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4381 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4441 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4501 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     4561 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     4621 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     4681 cttgatcaga gcttgatccc ctgcgccatc agatccttgg cggcgagaaa gccatccagt
     4741 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     4801 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     4861 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     4921 ggggtcagca ccgtttctgc ggactggctt tctacgtgaa aaggatctag gtgaagatcc
     4981 tttttgataa tctcatgcct gacatttata ttccccagaa catcaggtta atggcgtttt
     5041 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
     5101 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
     5161 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
     5221 gtcgccccgg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
     5281 ctcttttata ggtgtaaacc ttaaactgcc gtacgtatag gctgcgcaac tgttgggaag
     5341 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     5401 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     5461 gtgaattgta atacgactca ctatagggcg aattgggccc tctagatgca tgctcgagcg
     5521 gccgccagtg tgatggatat ctgcagaatt cgcccttga
//
