pSI-AGR10 vector (V003211)

LOCUS       40924_40197        5192 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-AGR10, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5192)
  AUTHORS   Mansouri M, Bellon-Echeverria I, Rizk A, Ehsaei Z, Cianciolo 
            Cosentino C, Silva CS, Xie Y, Boyce FM, Davis MW, Neuhauss SC, 
            Taylor V, Ballmer-Hofer K, Berger I, Berger P.
  TITLE     Highly efficient baculovirus-mediated multigene delivery in primary 
            cells
  JOURNAL   Nat Commun 7, 11529 (2016)
  PUBMED    27143231
REFERENCE   2  (bases 1 to 5192)
  AUTHORS   Mansouri M, Berger I, Berger P.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2016) Laboratory of Biomolecular Research, Paul 
            Scherrer Institute, OFLG-110, Villigen psi 5232, Switzerland
REFERENCE   3  (bases 1 to 5192)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5192)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            7, 11529 (2016)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2016) Laboratory of Biomolecular Research, Paul Scherrer 
            Institute, OFLG-110, Villigen psi 5232, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Ape v. v. v2.0.49
            Sequencing Technology :: 454; Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      190..778
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  complement(913..1136)
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1206..1736)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1925..1953)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2024..2057)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        2149..2452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2453..2656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2701..2719
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2781..3488
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     protein_bind    3547..3568
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3583..3613
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3621..3637
                     /label=lac operator
                     /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     3645..3661
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             3688..3710
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3835..4542
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     polyA_signal    4735..4959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     mobile_element  complement(5027..5192)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"