LOCUS       Exported                9816 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSV529-hTNF1, complete sequence.
ACCESSION   LT727561
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9816)
  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  JOURNAL   Unpublished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 9816)
  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 9816)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9816
                     /organism="Mammalian expression vector pSV529-hTNF1"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 2952."
                     /db_xref="taxon:1945114"
     promoter        82..439
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      101..382
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     rep_origin      290..425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      339..414
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             join(453..698,1045..2925)
                     /codon_start=1
                     /product="SV40 (simian virus 40) large T antigen"
                     /label=large T antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF
                     CSEEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT
                     TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI
                     CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK
                     CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA
                     VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP
                     KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN
                     FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR
                     TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL
                     LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD
                     DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     CDS             453..977
                     /codon_start=1
                     /note="unnamed protein product; SV40 small t-antigen"
                     /protein_id="SJL88843.1"
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEVFASSLNPGVDAMYCKQWPECA
                     KKMSANCICLLCLLRMKHENRKLYRKDPLVWVDCYCFDCFRMWFGLDLCEGTLLLWCDI
                     IGQTTYRDLKL"
     misc_feature    453..698
                     /label=SV40 large T-antigen
                     /note="SV40 large T-antigen"
     misc_feature    699..1044
                     /label=intron
                     /note="intron"
     intron          699..1044
                     /label=large T intron
                     /note="SV40 (simian virus 40) large T antigen intron"
     misc_feature    1045..2925
                     /label=SV40 large T-antigen
                     /note="SV40 large T-antigen"
     regulatory      complement(2926..3022)
                     /regulatory_class="terminator"
                     /note="polyA"
     regulatory      2926..3022
                     /regulatory_class="terminator"
                     /note="polyA"
     polyA_signal    2948..3082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(3085..3400)
                     /label=3UTR
                     /note="3UTR"
     CDS             complement(3401..3874)
                     /codon_start=1
                     /note="unnamed protein product; hTNF"
                     /protein_id="SJL88844.1"
                     /translation="VRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRD
                     NQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRE
                     TPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL"
     misc_feature    complement(3875..4102)
                     /label=phTNF
                     /note="phTNF"
     misc_feature    complement(4103..4280)
                     /label=5UTR
                     /note="5UTR"
     misc_feature    complement(4323..5260)
                     /label=intron
                     /note="intron"
     misc_feature    complement(5261..5463)
                     /label=5UTR
                     /note="5UTR"
     regulatory      complement(5464..5759)
                     /regulatory_class="promoter"
                     /note="SV40 late promoter"
     promoter        5496..5853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      5515..5796
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     rep_origin      5704..5839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      5753..5828
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             5867..6391
                     /codon_start=1
                     /note="unnamed protein product; SV40 small t-antigen"
                     /protein_id="SJL88845.1"
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEVFASSLNPGVDAMYCKQWPECA
                     KKMSANCICLLCLLRMKHENRKLYRKDPLVWVDCYCFDCFRMWFGLDLCEGTLLLWCDI
                     IGQTTYRDLKL"
     misc_feature    5867..6112
                     /label=SV40 large T-antigen
                     /note="SV40 large T-antigen"
     misc_feature    6113..6458
                     /label=intron
                     /note="intron"
     intron          6113..6458
                     /label=large T intron
                     /note="SV40 (simian virus 40) large T antigen intron"
     misc_feature    6459..7521
                     /label=part of SV40 large t-antigen
                     /note="part of SV40 large t-antigen"
     CDS             6588..6608
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    7663..7803
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7805..8207)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(7989..8577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8748..9608)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88846.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9609..9713)
                     /gene="bla"
                     /label=AmpR promoter