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
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 5LTR
                     /note="CRU5 chimeric CMV promoter, MoMLV 5LTR"
     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 5LTR
                     /note="R region 5LTR"
     misc_feature    670..745
                     /label=U5 region 5LTR
                     /note="U5 region 5LTR"
     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 3LTR, self-inactivating
                     /note="MoMLV 3LTR, self-inactivating"
     misc_feature    7058..7125
                     /label=R region 3LTR
                     /note="R region 3LTR"
     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