LOCUS       Exported                4146 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Donor vector pTKIP-cat, complete sequence.
ACCESSION   GU327536
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4146)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Site-specific chromosomal integration of large synthetic constructs
  JOURNAL   Nucleic Acids Res. 38 (6), E92 (2010)
  PUBMED    20047970
REFERENCE   2  (bases 1 to 4146)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2009) Molecular Biology, Princeton University, 
            Washington Rd., Princeton, NJ 08544, USA
REFERENCE   3  (bases 1 to 4146)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4146
                     /organism="Donor vector pTKIP-cat"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:711471"
     CDS             238..429
                     /codon_start=1
                     /gene="rop"
                     /product="ROP"
                     /label=rop
                     /protein_id="ADB98023.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            238..429
                     /gene="rop"
                     /label=rop
     misc_feature    531..671
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(857..1445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     ncRNA           complement(859..1410)
                     /label=antisense_RNA
                     /note="RNA II"
                     /ncRNA_class="antisense_RNA"
     ncRNA           1299..1406
                     /label=antisense_RNA
                     /note="RNA I"
                     /ncRNA_class="antisense_RNA"
     CDS             complement(1616..2476)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="ADB98022.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1616..2476)
                     /gene="bla"
                     /label=bla
     promoter        complement(2477..2581)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2607..2624
                     /label=I-SceI recognition site
                     /note="I-SceI recognition site"
     misc_feature    2625..2649
                     /label=Landing Pad Region 1
                     /note="Landing Pad Region 1"
     misc_feature    2656..2770
                     /note="multiple cloning site; MCS"
     primer_bind     2668..2684
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2706..2739
                     /label=FRT
                     /note="FRT"
     protein_bind    2706..2739
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      2797..3311
                     /regulatory_class="promoter"
                     /note="PGK promoter"
     promoter        2803..3302
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     regulatory      3314..3379
                     /regulatory_class="promoter"
                     /note="EM7 promoter"
     promoter        3314..3361
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3380..4039
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /function="chloramphenicol resistance"
                     /label=cat
                     /protein_id="ADB98021.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            3380..4039
                     /gene="cat"
                     /label=cat
     misc_feature    4070..4103
                     /label=FRT
                     /note="FRT"
     protein_bind    4070..4103
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    4104..4128
                     /label=Landing Pad Region 2
                     /note="Landing Pad Region 2"
     misc_feature    4129..4146
                     /label=I-SceI recognition site
                     /note="I-SceI recognition site"