pGBT-RP2.2 vector (V015520)

LOCUS       62056_11315        9118 bp DNA     circular SYN 25-AUG-2020
DEFINITION  Cloning vector pGBT-RP2.2, complete sequence.
ACCESSION   MT815588
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9118)
  AUTHORS   Ichino N, Serres MR, Urban RM, Urban MD, Treichel AJ, Schaefbauer 
            KJ, Greif LE, Varshney GK, Skuster KJ, McNulty MS, Daby CL, Wang Y, 
            Liao HK, El-Rass S, Ding Y, Liu W, Anderson JL, Wishman MD, 
            Sabharwal A, Schimmenti LA, Sivasubbu S, Balciunas D, Hammerschmidt 
            M, Farber SA, Wen XY, Xu X, McGrail M, Essner JJ, Burgess SM, Clark 
            KJ, Ekker SC.
  TITLE     Building the vertebrate codex using the gene breaking protein trap 
            library
  JOURNAL   Elife 9 (2020) In press
  PUBMED    32779569
REFERENCE   2  (bases 1 to 9118)
  AUTHORS   Ichino N, Serres MR, Urban RM, Urban MD, Treichel AJ, Schaefbauer 
            KJ, Greif LE, Varshney GK, Skuster KJ, McNulty MS, Daby CL, Wang Y, 
            Liao HK, El-Rass S, Ding Y, Liu W, Anderson JL, Wishman MD, 
            Sabharwal A, Schimmenti LA, Sivasubbu S, Balciunas D, Hammerschmidt 
            M, Farber SA, Wen XY, Xu X, McGrail M, Essner JJ, Burgess SM, Clark 
            KJ, Ekker SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUL-2020) Biochemistry and Molecular Biology, Mayo 
            Clinic, 200 First Street SW, Rochester, Olmsted, MN 55905, USA
REFERENCE   3  (bases 1 to 9118)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife 9 
            (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-JUL-2020) Biochemistry and Molecular Biology, Mayo Clinic, 200 
            First Street SW, Rochester, Olmsted, MN 55905, USA"
FEATURES             Location/Qualifiers
     source          1..9118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..43
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        58..88
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    96..112
                     /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     120..136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        154..172
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     repeat_region   222..413
                     /label=Tol2
                     /note="Tol2"
     misc_feature    430..447
                     /label=I-SceI
                     /note="I-SceI"
     misc_recomb     448..481
                     /label=loxP
                     /note="loxP"
     misc_feature    488..597
                     /note="derived from carp; contains beta actin splice
                     acceptor from intron1/exon2 boundary"
     CDS             652..1323
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     misc_feature    1339..2514
                     /note="contains ocean pout antifreeze poly(A) signal, 
                     terminator, putative border element"
     promoter        2576..2594
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2603..3738
                     /label=carp beta actin promoter region
                     /note="carp beta actin promoter region"
                     /regulatory_class="promoter"
     misc_feature    3744..3835
                     /note="derived from carp; contains beta actin non-coding
                     exon 1"
     misc_feature    3836..5099
                     /note="derived from carp; contains beta actin intron 1"
     CDS             5164..5877
                     /label=GFP
                     /note="green fluorescent protein"
     misc_feature    5944..6071
                     /note="derived from carp; contains beta actin splice-donor
                     from exon/intron 1 boundary"
     protein_bind    complement(6089..6122)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6121..6138
                     /label=I-SceI
                     /note="I-SceI"
     repeat_region   6139..6400
                     /label=Tol2
                     /note="Tol2"
     promoter        complement(6413..6431)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6438..6454)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6595..7050
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7128..7232
                     /label=AmpR promoter
     CDS             7233..8090
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8264..8852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"