LOCUS       Exported                9522 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pLV.hMyoD.eGFP, complete sequence.
ACCESSION   EU048697
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9522)
  AUTHORS   Goncalves MA, Swildens J, Holkers M, Narain A, van Nierop GP, van de
            Watering MJ, Knaan-Shanzer S, de Vries AA.
  TITLE     Genetic complementation of human muscle cells via directed stem cell
            fusion
  JOURNAL   Mol. Ther. 16 (4), 741-748 (2008)
  PUBMED    18334989
REFERENCE   2  (bases 1 to 9522)
  AUTHORS   Goncalves MA, de Vries AA.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JUL-2007) Molecular Cell Biology, Leiden University 
            Medical Center, Einthovenweg 20, Leiden 2333 ZC, The Netherlands
REFERENCE   3  (bases 1 to 9522)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9522
                     /organism="Shuttle vector pLV.hMyoD.eGFP"
                     /mol_type="other DNA"
                     /label=derived from Homo sapiens
                     /note="derived from Homo sapiens"
                     /db_xref="taxon:476185"
     repeat_region   1..411
                     /label=RSV-HIV-1 hybrid 5 LTR
                     /note="RSV-HIV-1 hybrid 5 LTR"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1799..1916
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      1930..2514
                     /regulatory_class="promoter"
                     /note="Homo sapiens cytomegalovirus immediate early gene 
                     promoter"
     enhancer        1939..2242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2243..2446
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2631..3593
                     /codon_start=1
                     /gene="MyoD1"
                     /product="MyoD1"
                     /label=MyoD1
                     /note="Homo sapiens myogenic transcription factor"
                     /protein_id="ABW34378.1"
                     /translation="MELLSPPLRDVDLTAPDGSLCSFATTDDFYDDPCFDSPDLRFFED
                     LDPRLMHVGALLKPEEHSHFPAAVHPAPGAREDEHVRAPSGHHQAGRCLLWACKACKRK
                     TTNADRRKAATMRERRRLSKVNEAFETLKRCTSSNPNQRLPKVEILRNAIRYIEGLQAL
                     LRDQDAAPPGAAAAFYAPGPLPPGRGGEHYSGDSDASSPRSNCSDGMMDYSGPPSGARR
                     RNCYEGAYYNEAPSEPRPGKSAAVSSLDCLSSIVERISTESPAAPALLLADVPSESPPR
                     RQEAAAPSEGESSGDPTQSPDAAPQCPAGANPNPIYQVL"
     gene            2631..3593
                     /gene="MyoD1"
                     /label=MyoD1
     misc_feature    3862..4436
                     /note="encephalomyocarditis virus internal ribosome entry 
                     site; IRES"
     misc_feature    3874..4426
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     regulatory      4441..4450
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4447..5166
                     /codon_start=1
                     /product="enhanced green fluorescent protein"
                     /label=enhanced green fluorescent protein
                     /note="reporter; eGFP"
                     /protein_id="ABW34379.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    4447..5163
                     /note="Homo sapiens hepatitis B post-transcriptional 
                     regulatory element (HPRE)"
     repeat_region   6037..6271
                     /label=HIV-1 SIN 3 LTR
                     /note="HIV-1 SIN 3 LTR"
     LTR             6037..6270
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6342..6463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6503..6638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6659..6677)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6687..6703)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6848..7303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7332..7436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7437..8297
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8468..9056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9344..9365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9380..9410
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9418..9434
                     /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)."
     primer_bind     9442..9458
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9479..9497
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"