LOCUS       40924_13465        6039 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-luc2-NEO, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  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  JOURNAL   Unpublished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 6039)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 6039)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6039)
  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            "Unpublished"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..6039
                     /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..2500
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    2504..2516
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(2598..2616)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2634..2650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2658..2674)
                     /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(2682..2712)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2727..2748)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3036..3624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3765..4136)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(4346..5137)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5489..5788)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5926..5942
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5949..5967
                     /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 gattgaacaa gatggattgc acgcaggttc
     1741 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     1801 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     1861 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     1921 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     1981 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2041 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2101 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     2161 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     2221 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     2281 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     2341 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     2401 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     2461 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaggcctct gtggcctcaa
     2521 gggcgaattc cagcacactg gcggccgtta ctagtggatc cgagctcggt accaagcttg
     2581 atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc atggtcatag
     2641 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2701 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2761 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2821 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2881 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2941 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3001 cccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3721 acctagatcc ttttaaatta aaaatgaagt tttagcacgt gtcagtcctg ctcctcggcc
     3781 acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc
     3841 tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc
     3901 gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc
     3961 ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg
     4021 gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc
     4081 gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt cgccatggtg
     4141 gccctcctca cgtgctatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4201 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4261 gtgccacctg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     4321 ggaaattgta agcgttaata attcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4381 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4441 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     4501 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4561 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4621 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     4681 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4741 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4801 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4861 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4921 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4981 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     5041 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5101 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     5161 cttgatcaga gcttgatccc ctgcgccatc agatccttgg cggcgagaaa gccatccagt
     5221 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     5281 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     5341 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     5401 ggggtcagca ccgtttctgc ggactggctt tctacgtgaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgcct gacatttata ttccccagaa catcaggtta atggcgtttt
     5521 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
     5581 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
     5641 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
     5701 gtcgccccgg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
     5761 ctcttttata ggtgtaaacc ttaaactgcc gtacgtatag gctgcgcaac tgttgggaag
     5821 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     5881 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     5941 gtgaattgta atacgactca ctatagggcg aattgggccc tctagatgca tgctcgagcg
     6001 gccgccagtg tgatggatat ctgcagaatt cgcccttga
//
