LOCUS       Exported               11506 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLvCmvLacZ.Gfp, complete sequence.
ACCESSION   EF205034
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11506)
  AUTHORS   van Tuyn J, Pijnappels DA, de Vries AA, de Vries I, van der 
            Velde-van Dijke I, Knaan-Shanzer S, van der Laarse A, Schalij MJ, 
            Atsma DE.
  TITLE     Fibroblasts from human postmyocardial infarction scars acquire 
            properties of cardiomyocytes after transduction with a recombinant 
            myocardin gene
  JOURNAL   FASEB J. 21 (12), 3369-3379 (2007)
  PUBMED    17579192
REFERENCE   2  (bases 1 to 11506)
  AUTHORS   van Tuyn J.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JAN-2007) MCB, LUMC, Einthovenweg 20, Leiden, ZH 2300 
            RC, Netherlands
REFERENCE   3  (bases 1 to 11506)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11506
                     /organism="Cloning vector pLvCmvLacZ.Gfp"
                     /mol_type="other DNA"
                     /db_xref="taxon:421868"
     protein_bind    546..567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        582..612
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    620..636
                     /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     644..660
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        681..699
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        727..953
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     repeat_region   936..1134
     LTR             954..1134
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    1181..1306
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1186..1325
                     /label=packaging signal
                     /note="packaging signal"
     misc_feature    1652..2509
                     /label=RRE
                     /note="RRE"
     misc_feature    1799..2032
                     /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    2523..2640
                     /label=CPPt
                     /note="CPPt"
     regulatory      2653..3169
                     /regulatory_class="promoter"
                     /note="CMV.IE"
     enhancer        2663..2966
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2967..3170
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3337..6393
                     /codon_start=1
                     /gene="LacZ"
                     /product="beta-galactosidase"
                     /label=LacZ
                     /protein_id="ABN09228.1"
                     /translation="MDPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQ
                     LRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITV
                     NPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLP
                     SEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHV
                     ATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYA
                     DRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLN
                     GKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDR
                     YGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESG
                     HGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKK
                     WLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDEN
                     GNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTS
                     EYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVR
                     VVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQF
                     NRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQ
                     AEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDT
                     PHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSEN
                     GLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIG
                     GDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     gene            3337..6393
                     /gene="LacZ"
                     /label=LacZ
     CDS             3346..6393
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     misc_feature    6556..7156
                     /label=ECMV IRES
                     /note="ECMV IRES"
     misc_feature    6584..7136
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     regulatory      7151..7160
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7157..7921
                     /codon_start=1
                     /gene="eGFP"
                     /product="enhanced green fluorescent protein"
                     /label=eGFP
                     /protein_id="ABN09229.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYTATTSISWPTSKDRHQG
                     ELQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQRPQREARSHGPAG
                     VRDRRRDHSRHGRAVQVKRPRRALWVPCMYSI"
     gene            7157..7921
                     /gene="eGFP"
                     /label=eGFP
     regulatory      7908..8663
                     /regulatory_class="other"
                     /note="HPRE"
     LTR             8745..8978
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     repeat_region   8779..8980
     polyA_signal    9050..9171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      9211..9346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(9367..9385)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9395..9411)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      9556..10011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        10040..10144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             10145..11005
                     /codon_start=1
                     /gene="AMPR"
                     /product="beta-lactamase"
                     /label=AMPR
                     /note="ampicillin resistance"
                     /protein_id="ABN09230.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            10145..11005
                     /gene="AMPR"
                     /label=AMPR
     rep_origin      11176..258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"