LOCUS       Exported                5947 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pU.CAG.Hand1, complete sequence.
ACCESSION   EF208956
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5947)
  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     Post MI scar fibroblasts can be differentiated into 
            cardiomyocyte-like cells using myocardin gene transfer
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5947)
  AUTHORS   van Tuyn J.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JAN-2007) MCB, LUMC, Einthovenweg 20, Leiden, ZH 2300 
            RC, Netherlands
REFERENCE   3  (bases 1 to 5947)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5947
                     /organism="Cloning vector pU.CAG.Hand1"
                     /mol_type="other DNA"
                     /db_xref="taxon:421898"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      68..1801
                     /regulatory_class="promoter"
                     /note="CAG"
     enhancer        68..447
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        450..725
                     /label=chicken beta-actin promoter
     intron          726..1734
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             1919..2566
                     /codon_start=1
                     /gene="HAND1"
                     /product="Hand1"
                     /label=HAND1
                     /note="eHand"
                     /protein_id="ABN09235.1"
                     /translation="MNLVGSYAHHHHHHHPHPAHPMLHEPFLFGPASRCHQERPYFQSW
                     LLSPADAAPDFPAGGPPPAAAAAATAYGPDARPGQSPGRLEALGGRLGRRKGSGPKKER
                     RRTESINSAFAELRECIPNVPADTKLSKIKTLRLATSYIAYLMDVLAKDAQSGDPEAFK
                     AELKKADGGRESKRKRELQQHEGFPPALGPVEKRIKGRTGWPQQVWALELNQ"
     gene            1919..2566
                     /gene="HAND1"
                     /label=HAND1
     CDS             1943..1963
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     misc_feature    2769..3295
                     /label=rabbit beta globin polyA signal
                     /note="rabbit beta globin polyA signal"
     polyA_site      2916..2971
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(3348..3364)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3838..3942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3943..4803
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGARERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4974..5562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5850..5871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5886..5916
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5924..5940
                     /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)."