LOCUS       Exported                8596 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pGL3-MSTN-3.8kb, complete sequence.
ACCESSION   JN542719
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8596)
  AUTHORS   Bi Y, Qiao X, Hua Z, Zhang L, Liu X, Li L, Hua W, Xiao H, Zhou J, 
            Wei Q, Zheng X.
  TITLE     An asymmetric PCR-based, reliable and rapid single-tube native DNA 
            engineering strategy
  JOURNAL   BMC Biotechnol. 12, 39 (2012)
  PUBMED    22768962
REFERENCE   2  (bases 1 to 8596)
  AUTHORS   Bi Y, Zheng X, Qiao X, Zhang R, Jiang L, Ouyang H, Shao C, Hua Z, 
            Zhou J, Liu X, Li L, Xiao H, Hua W, Zhang L, Wei Q.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-AUG-2011) Biotechnology Group, Institute of Animal 
            Science and Veterinary, Hubei Academy of Agricultural Science, No. 
            1, Nanhu Yaoyuan, Hongshan District, Wuhan, Hubei 430064, P. R. 
            China
REFERENCE   3  (bases 1 to 8596)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8596
                     /organism="Reporter vector pGL3-MSTN-3.8kb"
                     /mol_type="other DNA"
                     /db_xref="taxon:1219472"
     regulatory      61..3838
                     /regulatory_class="promoter"
                     /note="porcine MSTN promoter"
     regulatory      3707..3711
                     /regulatory_class="CAAT_signal"
     regulatory      3748..3754
                     /regulatory_class="TATA_box"
     regulatory      3772..3778
                     /regulatory_class="TATA_box"
     regulatory      3860..3869
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3866..5518
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=firefly luciferase
                     /protein_id="AFQ68242.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    5559..5680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6099..6687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6858..7718)
                     /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"
     promoter        complement(7719..7823)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7850..8305
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    8436..8484
                     /note="synthetic polyadenylation signal"
     misc_feature    8498..8589
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"