LOCUS       Exported                6683 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSGNluc, complete sequence.
ACCESSION   KY129798
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6683)
  AUTHORS   Kuri P, Ellwanger K, Kufer TA, Leptin M, Bajoghli B.
  TITLE     A high-sensitivity, bi-directional reporter to monitor NF-kappaB 
            activity in cell culture and zebrafish in real-time
  JOURNAL   J. Cell. Sci. (2016) In press
  PUBMED    27980067
REFERENCE   2  (bases 1 to 6683)
  AUTHORS   Bajoghli B.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2016) Directors Research, EMBL, Meyerhoferstrasse
            1, Heidelberg, BW 69117, Germany
REFERENCE   3  (bases 1 to 6683)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6683
                     /organism="Cloning vector pSGNluc"
                     /mol_type="other DNA"
                     /db_xref="taxon:1930210"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        649..667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(698..927)
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    703..910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(934..1653)
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=green fluorescent protein
                     /note="GFP"
                     /protein_id="APQ46081.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1650..1659
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      complement(1669..1928)
                     /regulatory_class="promoter"
                     /note="CMV minimal promoter"
     5UTR           complement(1674..1716)
                     /label=Xenopus globin 5-UTR
                     /note="translational enhancer from the 5-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     promoter        complement(1753..1771)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(1812..1830)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(1840..1858)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1884..1922)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     repeat_region   1945..2067
                     /rpt_family="8x NF-kappaB Binding Sequence"
                     /rpt_unit_seq="gggaattccc"
     regulatory      2099..2299
                     /regulatory_class="promoter"
                     /note="CMV minimal promoter"
     promoter        2105..2143
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        2169..2187
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2197..2215
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     5UTR           2252..2294
                     /label=Xenopus globin 5-UTR
                     /note="translational enhancer from the 5-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      2337..2346
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2343..3995
                     /codon_start=1
                     /product="luciferase"
                     /label=luciferase
                     /protein_id="APQ46082.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     5UTR           4001..4232
                     /label=beta globin
                     /note="beta globin"
     3UTR           4029..4154
                     /label=Xenopus globin 3-UTR
                     /note="3-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     regulatory      4233..4438
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    4236..4370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4474..4492)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4534..4550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4558..4574
                     /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(4582..4612)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4627..4648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4936..5524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5695..6555)
                     /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(6556..6660)
                     /gene="bla"
                     /label=AmpR promoter