LOCUS       40924_13455        5646 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-luc2-BSD, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5646)
  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 5646)
  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  JOURNAL   Unpublished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 5646)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   4  (bases 1 to 5646)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5646)
  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            "Unpublished"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..5646
                     /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..2107
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    2111..2123
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(2205..2223)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2241..2257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2265..2281)
                     /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(2289..2319)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2334..2355)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2643..3231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3372..3743)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(3953..4744)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5096..5395)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5533..5549
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5556..5574
                     /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 cctttgtctc aagaagaatc
     1741 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     1801 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     1861 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     1921 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     1981 cggacggtgt cgacaggtgc ttctcgatct gcatcctggg atcaaagcga tagtgaagga
     2041 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     2101 ggagggctaa ggcctctgtg gcctcaaggg cgaattccag cacactggcg gccgttacta
     2161 gtggatccga gctcggtacc aagcttgatg catagcttga gtattctata gtgtcaccta
     2221 aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2281 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2341 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2401 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2461 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2521 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2581 aacatgtgag caaaaggcca gcaaaagccc aggaaccgta aaaaggccgc gttgctggcg
     2641 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2701 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2761 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2821 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2881 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2941 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3001 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3061 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3121 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3181 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3241 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3301 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3361 agcacgtgtc agtcctgctc ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc
     3421 agggcgaact cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg
     3481 tcccggaagt tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc
     3541 acccacaccc aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac
     3601 agggtcacgt cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac
     3661 ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga
     3721 acggcactgg tcaacttcgc catggtggcc ctcctcacgt gctattattg aagcatttat
     3781 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3841 ggggttccgc gcacatttcc ccgaaaagtg ccacctgtat gcggtgtgaa ataccgcaca
     3901 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaataatt cagaagaact
     3961 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     4021 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     4081 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     4141 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     4201 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     4261 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     4321 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     4381 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     4441 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     4501 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     4561 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     4621 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     4681 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     4741 tcatgcgaaa cgatcctcat cctgtctctt gatcagagct tgatcccctg cgccatcaga
     4801 tccttggcgg cgagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg
     4861 gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc
     4921 gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc
     4981 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     5041 acgtgaaaag gatctaggtg aagatccttt ttgataatct catgcctgac atttatattc
     5101 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca
     5161 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc
     5221 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac
     5281 gtgcactggc cagggggatc accatccgtc gccccggcgt gtcaataata tcactctgta
     5341 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgccgta
     5401 cgtataggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5461 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     5521 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
     5581 tgggccctct agatgcatgc tcgagcggcc gccagtgtga tggatatctg cagaattcgc
     5641 ccttga
//
