LOCUS       40924_19816        4122 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with a kanamycin resistance marker, for mammalian 
            expression from a strongly compromised CMV promoter of a protein 
            with a cleavable C-terminal HaloTag(R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4122)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4122)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Subclone a coding sequence between the SgfI/AsiSI and 
            EcoICRI/Eco53kI sites to remove the lethal barnase gene.
FEATURES             Location/Qualifiers
     source          1..4122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..45
                     /label=CMVd3 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate
                     early promoter that yields strongly reduced expression 
                     relative to the full-length promoter"
     promoter        77..95
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        101..119
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             158..490
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     CDS             533..553
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="EDLYFQS"
     CDS             566..1453
                     /codon_start=1
                     /label=HaloTag(R)
                     /note="modified bacterial dehalogenase that forms covalent
                     bonds with chloroalkane derivatives"
                     /translation="SEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     polyA_signal    complement(1596..1717)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2116..2907
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(3082..3670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3786..4069
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
ORIGIN
        1 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
       61 cagatcacta gaagcttaat acgactcact atagggtacg atttaggtga cactatagaa
      121 taaggagcga tcgccatgga ataagtaagg aatccacatg gcacaggtta tcaacacgtt
      181 tgacggggtt gcggattatc ttcagacata tcataagcta cctgataatt acattacaaa
      241 atcagaagca caagccctcg gctgggtggc atcaaaaggg aaccttgcag acgtcgctcc
      301 ggggaaaagc atcggcggag acatcttctc aaacagggaa ggcaaactcc cgggcaaaag
      361 cggacgaaca tggcgtgaag cggatattaa ctatacatca ggcttcagaa attcagaccg
      421 gattctttac tcaagcgact ggctgattta caaaacaacg gaccattatc agacctttac
      481 aaaaatcaga taatgtttaa tgaccccgtg tcgagctctc gagccaacca ctgaggatct
      541 gtactttcag agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca
      601 ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac
      661 ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc
      721 gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga
      781 caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga
      841 agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt
      901 ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg
      961 ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg
     1021 caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct
     1081 gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt
     1141 cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg
     1201 tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc
     1261 tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc
     1321 tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct
     1381 gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct
     1441 ggagatttcc ggttaataga attctagagt cgacctgcag gcatgcaagc tgatccggct
     1501 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     1561 taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     1621 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1681 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1741 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     1801 atgtggtaaa atcgaattct aatggatcct ctttgcgctt gcgttttccc ttgtccagat
     1861 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     1921 gttccgcttc ctttagcagc ccttgcgccc tgagtgcttg cggcagcgtg agcttcaaaa
     1981 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg
     2041 gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga
     2101 tgacggtcgt ttcgcatgct tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2161 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2221 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2281 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     2341 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     2401 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     2461 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     2521 caagcgaaac atcgcatcga gcgagcacgc actcggatgg aagccggtct tgtcgatcag
     2581 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     2641 gcgcgtatgc cggatggtga ggatctcgtc gtgactcatg gcgatgcctg cttgccgaat
     2701 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     2761 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     2821 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     2881 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     2941 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3001 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3061 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3121 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3181 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3241 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3301 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3361 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3421 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3481 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3541 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3601 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3661 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3721 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3781 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     3841 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     3901 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     3961 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     4021 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     4081 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
