pSG-DV_AB vector (V015817)

Basic Vector Information

Vector Name:
pSG-DV_AB
Antibiotic Resistance:
Ampicillin
Length:
6846 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Stach C, McCann M, O'Brien C, Le TS
Promoter:
UbC

pSG-DV_AB vector Map

pSG-DV_AB6846 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600mutation to remove SapI sitelox2272FRT (minimal)BbsI siteGolden gate cloning scarAarI siteM13 fwdMCSM13 revlac operatorlac promoterCAP binding siteAarI siteGolden gate cloning scarBbsI siteUbC promoterBlasticidin-S deaminasepoly(A) signallox5171F3PhiC31 attPAmpRrrnB T1 terminatorrrnB T2 terminatororibom6xHisattB

pSG-DV_AB vector Sequence

LOCUS       V015817                 6846 bp    DNA     circular SYN 23-OCT-2019
DEFINITION  Exported.
ACCESSION   V015817
VERSION     V015817
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6846)
  AUTHORS   Stach C, McCann M, O'Brien C, Le TS, Somia N, Chen X, Lee K, Fu HY,
            Daoutidis P, Zhao L, Hu WS, Smanski MJ.
  TITLE     Model-driven engineering of N-linked glycosylation in Chinese
            Hamster Ovary cells
  JOURNAL   ACS Synth Biol (2019) In press
   PUBMED   31596566
REFERENCE   2  (bases 1 to 6846)
  AUTHORS   Stach CS.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-APR-2019) Biochemistry, Molecular Biology, and
            Biophysics; Chemical Engineering and Materials Science, University
            of Minnesota, 1479 Gortner Ave, Suite 344, Saint Paul, Minnesota
            55108, United States
REFERENCE   3  (bases 1 to 6846)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol (2019) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (30-APR-2019) Biochemistry, Molecular Biology, and Biophysics;
            Chemical Engineering and Materials Science, University of Minnesota,
            1479 Gortner Ave, Suite 344, Saint Paul, Minnesota 55108, United
            States"
FEATURES             Location/Qualifiers
     source          1..6846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    21..54
                     /label="lox2272"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
                     compatible with each other, but incompatible with loxP or
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(55..88)
                     /label="FRT (minimal)"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     misc_feature    89..94
                     /label="BbsI site"
                     /note="BbsI site"
     misc_feature    97..100
                     /label="Golden gate cloning scar"
                     /note="Golden gate cloning scar"
     misc_feature    105..111
                     /label="AarI site"
                     /note="AarI site"
     primer_bind     313..329
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    330..386
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(399..415)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(423..439)
                     /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)."
     promoter        complement(447..477)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(492..513)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    594..600
                     /label="AarI site"
                     /note="AarI site"
     misc_feature    605..608
                     /label="Golden gate cloning scar"
                     /note="Golden gate cloning scar"
     misc_feature    611..616
                     /label="BbsI site"
                     /note="BbsI site"
     promoter        622..1833
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             1865..2284
                     /gene="bsr"
                     /label="Blasticidin-S deaminase"
                     /note="Blasticidin-S deaminase from Bacillus cereus.
                     Accession#: P33967"
     polyA_signal    2334..2382
                     /label="poly(A) signal"
                     /note="synthetic polyadenylation signal"
     protein_bind    2383..2416
                     /label="lox5171"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are
                     compatible with each other, but incompatible with loxP or
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    2417..2450
                     /label="F3"
                     /note="F3"
     misc_feature    2469..2507
                     /label="PhiC31 attP"
                     /note="PhiC31 attP"
     misc_feature    2697
                     /label="mutation to remove SapI site"
                     /note="mutation to remove SapI site"
     CDS             complement(2833..3690)
                     /label="AmpR"
                     /note="beta-lactamase"
     terminator      4223..4309
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4401..4428
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      4562..5150
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    5330
                     /label="mutation to remove SapI site"
                     /note="mutation to remove SapI site"
     misc_feature    complement(5336..5476)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(6773..6790)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     protein_bind    6811..6844
                     /label="attB"
                     /note="minimal attB site for the phi-C31 integrase (Groth
                     et al., 2000)"

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