LOCUS       40924_19791        4182 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) vector with an ampicillin resistance marker, for mammalian 
            expression from a slightly compromised CMV promoter of a protein 
            with a C-terminal HaloTag(R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4182)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4182)
  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..4182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..100
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     promoter        132..150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        156..174
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             213..545
                     /codon_start=1
                     /label=barnase
                     /note="ribonuclease from Bacillus amyloliquefaciens"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     CDS             588..608
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="EDLYFQS"
     CDS             621..1508
                     /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(1651..1772)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2019..2123
                     /label=AmpR promoter
     CDS             2124..2981
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(3142..3730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3846..4129
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
ORIGIN
        1 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
       61 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      121 cactagaagc ttaatacgac tcactatagg gtacgattta ggtgacacta tagaataagg
      181 agcgatcgcc atggaataag taaggaatcc acatggcaca ggttatcaac acgtttgacg
      241 gggttgcgga ttatcttcag acatatcata agctacctga taattacatt acaaaatcag
      301 aagcacaagc cctcggctgg gtggcatcaa aagggaacct tgcagacgtc gctccgggga
      361 aaagcatcgg cggagacatc ttctcaaaca gggaaggcaa actcccgggc aaaagcggac
      421 gaacatggcg tgaagcggat attaactata catcaggctt cagaaattca gaccggattc
      481 tttactcaag cgactggctg atttacaaaa caacggacca ttatcagacc tttacaaaaa
      541 tcagataatg tttaatgacc ccgtgtcgag ctctcgagcc aaccactgag gatctgtact
      601 ttcagagcga taacgatgga tccgaaatcg gtactggctt tccattcgac ccccattatg
      661 tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg
      721 tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg
      781 ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac
      841 cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc
      901 tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact
      961 gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta
     1021 tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca
     1081 ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga
     1141 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     1201 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     1261 cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc
     1321 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc
     1381 tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc
     1441 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga
     1501 tttccggtta atagaattct agagtcgacc tgcaggcatg caagctgatc cggctgctaa
     1561 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1621 ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     1681 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1741 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1801 ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg
     1861 gtaaaatcga attctaatgg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc
     1921 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg
     1981 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2041 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2101 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2161 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2221 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2281 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc
     2341 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2401 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2461 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2521 ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2581 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2641 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2701 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2761 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2821 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2881 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2941 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc
     3001 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3061 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3121 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3181 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3241 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3301 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3361 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3421 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3481 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3541 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3601 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3661 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3721 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3781 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3841 atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg
     3901 aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat
     3961 aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt
     4021 tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa
     4081 ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca
     4141 gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
