LOCUS       40924_13490        5325 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-Rluc-NEO, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5325)
  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  JOURNAL   Unpublished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 5325)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5325)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5325)
  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            "Unpublished"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..5325
                     /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..1786
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    1790..1802
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(1884..1902)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1920..1936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1944..1960)
                     /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(1968..1998)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2013..2034)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2322..2910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3051..3422)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(3632..4423)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4775..5074)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5212..5228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5235..5253
                     /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 cggagggatt gaacaagatg gattgcacgc
     1021 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     1081 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     1141 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     1201 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     1261 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     1321 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     1381 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     1441 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     1501 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     1561 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     1621 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     1681 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     1741 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag gcctctgtgg
     1801 cctcaagggc gaattccagc acactggcgg ccgttactag tggatccgag ctcggtacca
     1861 agcttgatgc atagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg
     1921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2281 caaaagccca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2761 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3001 atcttcacct agatcctttt aaattaaaaa tgaagtttta gcacgtgtca gtcctgctcc
     3061 tcggccacga agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac
     3121 ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg
     3181 acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg
     3241 ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc
     3301 acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac
     3361 tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttcgcc
     3421 atggtggccc tcctcacgtg ctattattga agcatttatc agggttattg tctcatgagc
     3481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3541 cgaaaagtgc cacctgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3601 gcatcaggaa attgtaagcg ttaataattc agaagaactc gtcaagaagg cgatagaagg
     3661 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     3721 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     3781 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     3841 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     3901 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     3961 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4021 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4081 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4141 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4201 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4261 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4321 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4381 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     4441 ctgtctcttg atcagagctt gatcccctgc gccatcagat ccttggcggc gagaaagcca
     4501 tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg
     4561 gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag
     4621 ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     4681 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgaaaagg atctaggtga
     4741 agatcctttt tgataatctc atgcctgaca tttatattcc ccagaacatc aggttaatgg
     4801 cgtttttgat gtcattttcg cggtggctga gatcagccac ttcttccccg ataacggaga
     4861 ccggcacact ggccatatcg gtggtcatca tgcgccagct ttcatccccg atatgcacca
     4921 ccgggtaaag ttcacgggag actttatctg acagcagacg tgcactggcc agggggatca
     4981 ccatccgtcg ccccggcgtg tcaataatat cactctgtac atccacaaac agacgataac
     5041 ggctctctct tttataggtg taaaccttaa actgccgtac gtataggctg cgcaactgtt
     5101 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5161 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5221 cggccagtga attgtaatac gactcactat agggcgaatt gggccctcta gatgcatgct
     5281 cgagcggccg ccagtgtgat ggatatctgc agaattcgcc cttga
//
