LOCUS       40924_13460        6273 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-luc2-HYG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6273)
  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  JOURNAL   Unpublished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 6273)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 6273)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6273)
  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            "Unpublished"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..6273
                     /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..2734
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    2738..2750
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(2832..2850)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2868..2884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2892..2908)
                     /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(2916..2946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2961..2982)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3270..3858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3999..4370)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(4580..5371)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5723..6022)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6160..6176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6183..6201
                     /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 gatgaaaaag cctgaactca ccgcgacgtc
     1741 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     1801 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     1861 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     1921 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg
     1981 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     2041 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     2101 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     2161 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     2221 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     2281 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     2341 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     2401 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     2461 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     2521 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     2581 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     2641 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     2701 ccgacgcccc agcactcgtc cgagggcaaa ggaatagggc ctctgtggcc tcaagggcga
     2761 attccagcac actggcggcc gttactagtg gatccgagct cggtaccaag cttgatgcat
     2821 agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt
     2881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3181 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaagcccagg
     3241 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3721 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3781 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3841 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3901 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3961 atccttttaa attaaaaatg aagttttagc acgtgtcagt cctgctcctc ggccacgaag
     4021 tgcacgcagt tgccggccgg gtcgcgcagg gcgaactccc gcccccacgg ctgctcgccg
     4081 atctcggtca tggccggccc ggaggcgtcc cggaagttcg tggacacgac ctccgaccac
     4141 tcggcgtaca gctcgtccag gccgcgcacc cacacccagg ccagggtgtt gtccggcacc
     4201 acctggtcct ggaccgcgct gatgaacagg gtcacgtcgt cccggaccac accggcgaag
     4261 tcgtcctcca cgaagtcccg ggagaacccg agccggtcgg tccagaactc gaccgctccg
     4321 gcgacgtcgc gcgcggtgag caccggaacg gcactggtca acttcgccat ggtggccctc
     4381 ctcacgtgct attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4501 cctgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     4561 tgtaagcgtt aataattcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     4621 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     4681 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     4741 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4801 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     4861 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     4921 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4981 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     5041 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     5101 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     5161 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     5221 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     5281 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     5341 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     5401 cagagcttga tcccctgcgc catcagatcc ttggcggcga gaaagccatc cagtttactt
     5461 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     5521 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
     5581 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     5641 agcaccgttt ctgcggactg gctttctacg tgaaaaggat ctaggtgaag atcctttttg
     5701 ataatctcat gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
     5761 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
     5821 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
     5881 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
     5941 ccggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
     6001 tataggtgta aaccttaaac tgccgtacgt ataggctgcg caactgttgg gaagggcgat
     6061 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6121 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     6181 tgtaatacga ctcactatag ggcgaattgg gccctctaga tgcatgctcg agcggccgcc
     6241 agtgtgatgg atatctgcag aattcgccct tga
//
