LOCUS       40924_9281        8276 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCD-A7M8K-p20mut, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8276)
  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  JOURNAL   Unpublished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 8276)
  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 8276)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8276)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      927..936
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             933..2256
                     /codon_start=1
                     /note="unnamed protein product; hAPI2 fragment"
                     /protein_id="SJL86949.1"
                     /translation="MDIVENSIFLSNLMKSANTFELKYDLSCELYRMSTYSTFPAGVPV
                     SERSLARAGFYYTGVNDKVKCFCCGLMLDNWKRGDSPTEKHKKLYPSCRFVQSLNSVNN
                     LEATSQPTFPSSVTNSTHSLLPGTENSGYFRGSYSNSPSNPVNSRANQDFSALMRSSYH
                     CAMNNENARLLTFQTWPLTFLSPTDLAKAGFYYIGPGDRVACFACGGKLSNWEPKDNAM
                     SEHLRHFPKCPFIENQLQDTSRYTVSNLSMQTHAARFKTFFNWPSSVLVNPEQLASAGF
                     YYVGNSDDVKCFCCDGGLRCWESGDDPWVQHAKWFPRCEYLIRIKGQEFIRQVQASYPH
                     LLEQLLSTSDSPGDENAESSIIHFEPGEDHSEDAIMMNTPVINAAVEMGFSRSLVKQTV
                     QRKILATGENYRLVNDLVLDLLNAEDEIREEERERATEEKES"
     misc_feature    936..938
                     /label=Kozak
                     /note="Kozak"
     misc_feature    2184..2186
                     /label=PCR artefact
                     /note="PCR artefact"
     CDS             2257..3770
                     /codon_start=1
                     /note="unnamed protein product; hMLT mutated fragment"
                     /protein_id="SJL86950.1"
                     /translation="MN*IILVILIIKSKQLTSLWRRTRLPF**EI*ITGSTPSSKLLWW
                     MCTN*LTY*DSWTSKWFHCWILLNMRCVMLWMSFYSF*TREYMGYYIMQDMVMKILGTA
                     SWSPLMLQIHIGLKIVCVYKIY*N*CKKKKLDLMCSYWIWRAKEMTTMIPFQSWMH*KS
                     PPILCLDMPRVKELKLLKSSILDWQMESL*NF*KTDY*KIRKSLCYWMKLQKIWVSVTL
                     PKANRL*RFEVVYLRREHLLIQYREQNILLNLLCGIYSGPRLMNFQKVCVLSLTVVFRF
                     N*DLQLSFPMS*SSIQV*FTNHRR**CVMPTLLIFHLI*ILIQKMQIKARLKKLAATWY
                     QRIFPSIASIPDSVHCKN*RNI*SSQYVYHISTQDWKIL*RTSRK*MLGNLSLLN*TCI
                     EVWEGRLAFKLVLCLMVLTRVLQPPQEEQGIITHCKTHSMVFTIHILVIQVMLHQQIAV
                     IAAGLQMHLFQVSLTMLHVILVEVMCQ*RQLMKYHLVSLTGSEFLK"
     misc_feature    2688..2693
                     /label=C464A
                     /note="C464A"
     misc_feature    2784..2786
                     /label=stability creator
                     /note="stability creator"
     misc_feature    3246..3248
                     /label=PCR artefact
                     /note="PCR artefact"
     CDS             3780..3809
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             3825..3842
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    3871..4095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4141..4569
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4583..4912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4979..5770
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5949..6082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6119..6135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6143..6159)
                     /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(6167..6197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6212..6233)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6521..7109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7283..8140)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8141..8245)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatcctgcca ccatggacat agtagaaaac agcatattct
      961 tatcaaattt gatgaaaagc gccaacacgt ttgaactgaa atacgacttg tcatgtgaac
     1021 tgtaccgaat gtctacgtat tccacttttc ctgctggggt tcctgtctca gaaaggagtc
     1081 ttgctcgtgc tggtttctat tacactggtg tgaatgacaa ggtcaaatgc ttctgttgtg
     1141 gcctgatgct ggataactgg aaaagaggag acagtcctac tgaaaagcat aaaaagttgt
     1201 atcctagctg cagattcgtt cagagtctaa attccgttaa caacttggaa gctacctctc
     1261 agcctacttt tccttcttca gtaacaaatt ccacacactc attacttccg ggtacagaaa
     1321 acagtggata tttccgtggc tcttattcaa actctccatc aaatcctgta aactccagag
     1381 caaatcaaga tttttctgcc ttgatgagaa gttcctacca ctgtgcaatg aataacgaaa
     1441 atgccagatt acttactttt cagacatggc cattgacttt tctgtcgcca acagatctgg
     1501 caaaagcagg cttttactac ataggacctg gagacagagt ggcttgcttt gcctgtggtg
     1561 gaaaattgag caattgggaa ccgaaggata atgctatgtc agaacacctg agacattttc
     1621 ccaaatgccc atttatagaa aatcagcttc aagacacttc aagatacaca gtttctaatc
     1681 tgagcatgca gacacatgca gcccgcttta aaacattctt taactggccc tctagtgttc
     1741 tagttaatcc tgagcagctt gcaagtgcgg gtttttatta tgtgggtaac agtgatgatg
     1801 tcaaatgctt ttgctgtgat ggtggactca ggtgttggga atctggagat gatccatggg
     1861 ttcaacatgc caagtggttt ccaaggtgtg agtacttgat aagaattaaa ggacaggagt
     1921 tcatccgtca agttcaagcc agttaccctc atctacttga acagctgcta tccacatcag
     1981 acagcccagg agatgaaaat gcagagtcat caattatcca ttttgaacct ggagaagacc
     2041 attcagaaga tgcaatcatg atgaatactc ctgtgattaa tgctgccgtg gaaatgggct
     2101 ttagtagaag cctggtaaaa cagacagttc agagaaaaat cctagcaact ggagagaatt
     2161 atagactagt caatgatctt gtgttggact tactcaatgc agaagatgaa ataagggaag
     2221 aggagagaga aagagcaact gaggaaaaag aatcaaatga attaaataat cttggtcatc
     2281 ctgataataa agagcaaaca actgaccagc ctttggcgaa ggacaaggtt gcccttttga
     2341 taggaaatat gaattaccgg gagcacccca agctcaaagc tcctttggtg gatgtgtacg
     2401 aattgactaa cttactgaga cagctggact tcaaagtggt ttcactgttg gatcttactg
     2461 aatatgagat gcgtaatgct gtggatgagt ttttactcct tttagacaag ggagtatatg
     2521 ggttattata ttatgcagga catggttatg aaaattttgg gaacagcttc atggtccccg
     2581 ttgatgctcc aaatccatat aggtctgaaa attgtctgtg tgtacaaaat atactgaaat
     2641 tgatgcaaga aaaagaaact ggacttaatg tgttcttatt ggatatggcg cgcaaaagaa
     2701 atgactacga tgataccatt ccaatcttgg atgcactaaa agtcaccgcc aatattgtgt
     2761 ttggatatgc cacgtgtcaa ggagctgaag cttttgaaat ccagcattct ggattggcaa
     2821 atggaatctt tatgaaattt ttaaaagaca gattattaga agataagaaa atcactgtgt
     2881 tactggatga agttgcagaa gatatgggta agtgtcacct taccaaaggc aaacaggctc
     2941 tagagattcg aagtagttta tctgagaaga gagcacttac tgatccaata cagggaacag
     3001 aatattctgc tgaatctctt gtgcggaatc tacagtgggc caaggctcat gaacttccag
     3061 aaagtatgtg tcttaagttt gactgtggtg ttcagattca attaggattt gcagctgagt
     3121 tttccaatgt catgatcatc tatacaagta tagtttacaa accaccggag ataataatgt
     3181 gtgatgccta cgttactgat tttccacttg atctagatat tgatccaaaa gatgcaaata
     3241 aaggcacgcc tgaagaaact ggcagctact tggtatcaaa ggatcttccc aagcattgcc
     3301 tctataccag actcagttca ctgcaaaaat taaaggaaca tctagtcttc acagtatgtt
     3361 tatcatatca gtactcagga ttggaagata ctgtagagga caagcaggaa gtgaatgttg
     3421 ggaaacctct cattgctaaa ttagacatgc atcgaggttt gggaaggaag acttgctttc
     3481 aaacttgtct tatgtctaat ggtccttacc agagttctgc agccacctca ggaggagcag
     3541 ggcattatca ctcattgcaa gacccattcc atggtgttta ccattcacat cctggtaatc
     3601 caagtaatgt tacaccagca gatagctgtc attgcagccg gactccagat gcatttattt
     3661 caagtttcgc tcaccatgct tcatgtcatt ttagtagaag taatgtgcca gtagagacaa
     3721 ctgatgaaat accatttagt ttctctgaca ggctcagaat ttctgaaaaa gggcccttcg
     3781 aacaaaaact catctcagaa gaggatctga atatgcatac cggtcatcat caccatcacc
     3841 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3901 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3961 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4021 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     4081 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     4141 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4201 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4261 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4321 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4381 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4441 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4501 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4561 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4621 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4681 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4741 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4801 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4861 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4921 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4981 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5041 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5101 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5161 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5221 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5281 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5341 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5401 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5461 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5521 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5581 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5641 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5701 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5761 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     5821 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5881 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5941 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6001 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6061 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6121 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6181 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6241 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6301 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6961 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7201 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7261 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7381 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7441 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7561 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8161 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8221 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
