LOCUS       40924_13480        4932 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRm-Rluc-BSD, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4932)
  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  JOURNAL   Unpublished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 4932)
  AUTHORS   Yates PA, Soysa R, Carter NS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 4932)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4932)
  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            "Unpublished"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..4932
                     /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..1393
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    1397..1409
                     /label=SfiI restriction site
                     /note="SfiI restriction site"
     promoter        complement(1491..1509)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1527..1543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1551..1567)
                     /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(1575..1605)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1620..1641)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1929..2517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2658..3029)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             complement(3239..4030)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4382..4681)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4819..4835
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4842..4860
                     /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 gccaagcctt tgtctcaaga
     1021 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     1081 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     1141 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     1201 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     1261 cccctgcgga cggtgtcgac aggtgcttct cgatctgcat cctgggatca aagcgatagt
     1321 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     1381 tgtgtgggag ggctaaggcc tctgtggcct caagggcgaa ttccagcaca ctggcggccg
     1441 ttactagtgg atccgagctc ggtaccaagc ttgatgcata gcttgagtat tctatagtgt
     1501 cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1561 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1621 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1681 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1741 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1801 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1861 ggaaagaaca tgtgagcaaa aggccagcaa aagcccagga accgtaaaaa ggccgcgttg
     1921 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1981 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2041 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2101 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2161 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2221 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2281 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2341 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     2401 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2461 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2521 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2581 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2641 agttttagca cgtgtcagtc ctgctcctcg gccacgaagt gcacgcagtt gccggccggg
     2701 tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat ggccggcccg
     2761 gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag ctcgtccagg
     2821 ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg gaccgcgctg
     2881 atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac gaagtcccgg
     2941 gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg cgcggtgagc
     3001 accggaacgg cactggtcaa cttcgccatg gtggccctcc tcacgtgcta ttattgaagc
     3061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgtatgcgg tgtgaaatac
     3181 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta ataattcaga
     3241 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     3301 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     3361 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     3421 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     3481 gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     3541 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     3601 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     3661 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     3721 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     3781 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     3841 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     3901 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     3961 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     4021 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agagcttgat cccctgcgcc
     4081 atcagatcct tggcggcgag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     4141 cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta
     4201 gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc
     4261 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg
     4321 ctttctacgt gaaaaggatc taggtgaaga tcctttttga taatctcatg cctgacattt
     4381 atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat
     4441 cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc
     4501 gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca
     4561 gcagacgtgc actggccagg gggatcacca tccgtcgccc cggcgtgtca ataatatcac
     4621 tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact
     4681 gccgtacgta taggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4741 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4801 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
     4861 gcgaattggg ccctctagat gcatgctcga gcggccgcca gtgtgatgga tatctgcaga
     4921 attcgccctt ga
//
