LOCUS       40924_13495        5133 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-Rluc-PAC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5133)
  AUTHORS   Martin JL, Yates PA, Soysa R, Alfaro JF, Yang F, Burnum-Johnson KE, 
            Petyuk VA, Weitz KK, Camp DG II, Smith RD, Wilmarth PA, David LL, 
            Ramasamy G, Myler PJ, Carter NS.
  TITLE     Metabolic Reprogramming during Purine Stress in the Protozoan 
            Pathogen Leishmania donovani
  JOURNAL   PLoS Pathog. 10 (2), E1003938 (2014)
  PUBMED    24586154
REFERENCE   2  (bases 1 to 5133)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     A dual luciferase system for analysis of post-transcriptional gene 
            expression in Leishmania
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5133)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   4  (bases 1 to 5133)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5133)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2014"; volume: "10"; issue: "2"; pages: "E1003938"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (13-MAY-2013) Biochemistry "
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5133
                     /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..1594
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    1599..1610
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(1692..1710)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1728..1744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1752..1768)
                     /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(1776..1806)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1821..1842)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2130..2718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2859..3230)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(3440..4231)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4583..4882)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5020..5036
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5043..5061
                     /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 accgagtaca agcccacggt
     1021 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     1081 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     1141 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     1201 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     1261 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     1321 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     1381 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     1441 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     1501 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     1561 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaggc ctctgtggcc tcaagggcga
     1621 attccagcac actggcggcc gttactagtg gatccgagct cggtaccaag cttgatgcat
     1681 agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt
     1741 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1801 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1861 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1921 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1981 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2041 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaagcccagg
     2101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2821 atccttttaa attaaaaatg aagttttagc acgtgtcagt cctgctcctc ggccacgaag
     2881 tgcacgcagt tgccggccgg gtcgcgcagg gcgaactccc gcccccacgg ctgctcgccg
     2941 atctcggtca tggccggccc ggaggcgtcc cggaagttcg tggacacgac ctccgaccac
     3001 tcggcgtaca gctcgtccag gccgcgcacc cacacccagg ccagggtgtt gtccggcacc
     3061 acctggtcct ggaccgcgct gatgaacagg gtcacgtcgt cccggaccac accggcgaag
     3121 tcgtcctcca cgaagtcccg ggagaacccg agccggtcgg tccagaactc gaccgctccg
     3181 gcgacgtcgc gcgcggtgag caccggaacg gcactggtca acttcgccat ggtggccctc
     3241 ctcacgtgct attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3301 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3361 cctgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     3421 tgtaagcgtt aataattcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     3481 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     3541 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     3601 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     3661 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     3721 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     3781 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     3841 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     3901 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     3961 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     4021 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     4081 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     4141 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4201 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     4261 cagagcttga tcccctgcgc catcagatcc ttggcggcga gaaagccatc cagtttactt
     4321 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     4381 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
     4441 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     4501 agcaccgttt ctgcggactg gctttctacg tgaaaaggat ctaggtgaag atcctttttg
     4561 ataatctcat gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
     4621 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
     4681 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
     4741 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
     4801 ccggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
     4861 tataggtgta aaccttaaac tgccgtacgt ataggctgcg caactgttgg gaagggcgat
     4921 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     4981 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     5041 tgtaatacga ctcactatag ggcgaattgg gccctctaga tgcatgctcg agcggccgcc
     5101 agtgtgatgg atatctgcag aattcgccct tga
//
