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L149 vector

Basic Vector Information

Basic Vector Information
Vector Name L149 Antibiotic Resistance Ampicillin
Length 9502 bp Type Retroviral expression vector
Source Micklem DR, Lorens JB.

L149 vector Vector Map

L149 vector Sequence

Copy Sequence

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LOCUS       Exported                9502 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Retroviral expression vector L149, complete sequence.
ACCESSION   EU753858
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9502)
  AUTHORS   Micklem DR, Lorens JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAY-2008) Department of Biomedicine, University of 
            Bergen, Jonas Lies vei 91, Bergen 5009, Norway
REFERENCE   2  (bases 1 to 9502)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 18, 2018 from SnapGene 4.1.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9502
                     /organism="Retroviral expression vector L149"
                     /mol_type="other DNA"
                     /db_xref="taxon:563906"
     repeat_region   1..745
                     /label=CRU5 chimeric CMV promoter, MoMLV 5'LTR
                     /note="CRU5 chimeric CMV promoter, MoMLV 5'LTR"
     regulatory      1..570
                     /regulatory_class="enhancer"
                     /note="CMV immediate-early enhancer region"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    601..609
                     /label=R region 5'LTR
                     /note="R region 5'LTR"
     misc_feature    670..745
                     /label=U5 region 5'LTR
                     /note="U5 region 5'LTR"
     misc_feature    745..1655
                     /label=MoMLV extended packaging signal (psi+)
                     /note="MoMLV extended packaging signal (psi+)"
     stem_loop       804..828
                     /label=retroviral packaging stem-loop A
                     /note="retroviral packaging stem-loop A"
     misc_feature    808..1165
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     stem_loop       878..909
                     /label=retroviral packaging stem-loop B
                     /note="retroviral packaging stem-loop B"
     stem_loop       910..952
                     /label=retroviral packaging stem-loop C
                     /note="retroviral packaging stem-loop C"
     stem_loop       955..974
                     /label=retroviral packaging stem-loop D
                     /note="retroviral packaging stem-loop D"
     CDS             1230..1646
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     misc_feature    1656..2030
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     misc_feature    complement(2048..2256)
                     /note="BGHpA signal; bovine growth hormone polyadenylation 
                     signal"
     polyA_signal    2049..2256
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     mRNA            complement(join(2143..6229,6363..6464))
                     /gene="P2AG2ALuc2ANTR"
                     /product="Puromycin2AGFP2ALuciferase2ANTR"
                     /label=P2AG2ALuc2ANTR mRNA
     gene            complement(2143..6464)
                     /gene="P2AG2ALuc2ANTR"
                     /label=P2AG2ALuc2ANTR
     polyA_site      complement(2143)
                     /gene="P2AG2ALuc2ANTR"
     CDS             complement(2292..6146)
                     /codon_start=1
                     /gene="P2AG2ALuc2ANTR"
                     /product="Puromycin2AGFP2ALuciferase2ANTR"
                     /label=P2AG2ALuc2ANTR
                     /note="confers puromycin resistance, green fluorescence and
                     nitroreductase activity; no luciferase activity detected"
                     /protein_id="ACI23257.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAGSGQLLNFDLLKLAGDVESNPGP
                     MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWP
                     TLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGD
                     TLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSV
                     QLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE
                     LYKEFSGLRSGQLLNFDLLKLAGDVESNPGPMEDAKNIKKGPAPFYPLEDGTAGEQLHK
                     AMKRYALVPGTIAFTDAHIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQ
                     FFMPVLGALFIGVAVAPANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPII
                     QKIIIMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLP
                     KGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVL
                     MYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEV
                     GEAVAKRFHLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTL
                     GVNQRGELCVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSL
                     IKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVD
                     YVASQVTTAKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKIAVEFSGLRSGQ
                     LLNFDLLKLAGDVESNPGPMDIISVALKRHSTKAFDASKKLTPEQAEQIKTLLQYSPSS
                     TNSQPWHFIVASTEEGKARVAKSAAGNYVFNERKMLDASHVVVFCAKTAMDDVWLKLVV
                     DQEDADGRFATPEAKAANDKGRKFFADMHRKDLHDDAEWMAKQVYLNVGNFLLGVAALG
                     LDAVPIEGFDAAILDAEFGLKEKGYTSLVVVPVGHHSVEDFNATLPKSRLPQNITLTEV
                     "
     misc_feature    complement(2946..3029)
                     /gene="P2AG2ALuc2ANTR"
                     /label=FMDV 2A self-cleaving linker region
                     /note="FMDV 2A self-cleaving linker region"
     CDS             complement(3030..4679)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    complement(4680..4763)
                     /gene="P2AG2ALuc2ANTR"
                     /label=FMDV 2A self-cleaving linker region
                     /note="FMDV 2A self-cleaving linker region"
     CDS             complement(4764..5480)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    5481..5549
                     /label=FMDV 2A self-cleaving linker region
                     /note="FMDV 2A self-cleaving linker region"
     CDS             complement(5553..6146)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPG"
     intron          6230..6362
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      complement(6465..6517)
                     /regulatory_class="promoter"
                     /note="PhCMV* minimal promoter"
     promoter        complement(6480..6517)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_binding    complement(6535..6833)
                     /label=Tet-activator/repressor binding site
                     /bound_moiety="Tet-activator/repressor"
                     /note="Tet operator (seven copies)"
     protein_bind    6550..6820
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     misc_feature    6892..7056
                     /label=U3SIN (self-inactivating) region
                     /note="U3SIN (self-inactivating) region"
     repeat_region   6892..7056
                     /label=MoMLV 3'LTR, self-inactivating
                     /note="MoMLV 3'LTR, self-inactivating"
     misc_feature    7058..7125
                     /label=R region 3'LTR
                     /note="R region 3'LTR"
     primer_bind     complement(7278..7294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7302..7318
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(7326..7356)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7371..7392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7665..8377)
                     /direction=LEFT
                     /label=pMB1 ori (pUC)
                     /note="pMB1 ori (pUC)"
     rep_origin      complement(7680..8268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8442..9302)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ACI23256.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     mRNA            complement(8442..9302)
                     /gene="bla"
                     /label=bla mRNA
     gene            complement(8442..9302)
                     /gene="bla"
                     /label=bla
     promoter        complement(9303..9407)
                     /gene="bla"
                     /label=AmpR promoter

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