LOCUS       Exported                6213 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAMFhGNTIf1, complete sequence.
ACCESSION   LT727444
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6213)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6213)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 6213)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6213
                     /organism="Mammalian expression vector pCAMFhGNTIf1"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3124."
                     /db_xref="taxon:1944901"
     regulatory      82..173
                     /regulatory_class="terminator"
                     /note="polyA"
     polyA_site      149..204
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    174..530
                     /label=3FR
                     /note="3FR"
     primer_bind     complement(565..581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    589..605
                     /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)."
     regulatory      complement(608..690)
                     /regulatory_class="promoter"
                     /note="lac"
     promoter        complement(613..643)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    657..678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        737..932
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      783..918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      832..907
                     /label=SV40 ori
                     /note="SV40 ori"
     polyA_signal    938..1072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(938..1012)
                     /regulatory_class="terminator"
                     /note="polyA"
     regulatory      938..1012
                     /regulatory_class="terminator"
                     /note="polyA"
     rep_origin      complement(1124..1528)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(1310..1898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2069..2929)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88471.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2930..3034)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3065..3444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      3065..3442
                     /regulatory_class="promoter"
                     /note="hCMV"
     promoter        3446..3723
                     /label=chicken beta-actin promoter
     regulatory      3448..4696
                     /regulatory_class="promoter"
                     /note="AG"
     misc_feature    3817..4696
                     /label=intron
                     /note="intron"
     CDS             4707..4958
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=-alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDK"
     misc_feature    4707..4949
                     /label=ppMF
                     /note="ppMF"
     CDS             4962..6176
                     /codon_start=1
                     /note="unnamed protein product; hGNTI"
                     /protein_id="SJL88472.1"
                     /translation="ALDGDPASLTREVIRLAQDAEVELERQRGLLQQIGDALSSQRGRV
                     PTAAPPAQPRVPVTPAPAVIPILVIACDRSTVRRCLDKLLHYRPSAELFPIIVSQDCGH
                     EETAQAIASYGSAVTHIRQPDLSSIAVPPDHRKFQGYYKIARHYRWALGQVFRQFRFPA
                     AVVVEDDLEVAPDFFEYFRATYPLLKADPSLWCVSAWNDNGKEQMVDASRPELLYRTDF
                     FPGLGWLLLAELWAELEPKWPKAFWDDWMRRPEQRQGRACIRPEISRTMTFGRKGVSHG
                     QFFDQHLKFIKLNQQFVHFTQLDLSYLQREAYDRDFLARVYGAPQLQVEKVRTNDRKEL
                     GEVRVQYTGRDSFKAFAKALGVMDDLKSGVPRAGYRGIVTFQFRGRRVHLAPPLTWEGY
                     DPSWN"