LOCUS       Exported                8014 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses pentamerised full-length protein ectodomain with no 
            N-linked glycans in mammalian cells. Contains a C-terminal rat Cd4 
            d3+4 tag, COMP pentamerisation domain, flag- and 6xHis tags. .
ACCESSION   .
VERSION     .
KEYWORDS    PF3D7_1310500-COMP-blac-flag-his
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8014)
  TITLE     Pentamerised full-length protein ectodomains with no N-linked 
            glycans
REFERENCE   2  (bases 1 to 8014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8014
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     310..329
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      352..361
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1546..2331
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     CDS             2338..2403
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2410..2427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2465..2520
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2519..2538)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4756..4774)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4842..4946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4947..5807
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5978..6566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6467..6486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6720..6737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6854..6875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6890..6920
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6928..6944
                     /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)."
     primer_bind     6933..6955
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6952..6968
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6952..6968
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7239..7618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7619..7822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7773..7793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7819..7843
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ctcgagaaag gcgtctaacc agtcacagtc gcaaggtagg ctgagcaccg tggcgggcgg
       61 cagcgggtgg cggtcggggt tgtttctggc ggaggtgctg ctgatgatgt aattaaagta
      121 ggcggtcttg agacggcgga tggtcgaggt gaggtgtggc aggcttgaga tccagctgtt
      181 ggggtgagta ctccctctca aaagcgggca ttacttctgc gctaagattg tcagtttcca
      241 aaaacgagga ggatttgata ttcacctggc ccgatctggc catacacttg agtgacaatg
      301 acatccactt tgcctttctc tccacaggtg tccactccca ggtccaagtt tgccaccatg
      361 gagtttcaga cccaggtact catgtccctg ctgctctgca tgtctggtgc ggccgccgat
      421 gaggacgtga tcgagctgaa gaacaactac atcgagggca tcgtgaaggg cgacgccgac
      481 ctgctgttca actgcgccat ctacctggac agcgacacct acgtgtcccc aaagagcgac
      541 ggcaagttcc tgttcaagaa cgtgaaagag ggcgagtaca ctatgtacgt gaaccacccc
      601 tacctcgagt tcaacaagta ctacgtggaa gtgaagaagc agatcacccg gaacaacggc
      661 gccctgtacc tggtgcaggc ctacgagctg aacctgccct tcgagaagtc caacctgatc
      721 gtgaacaaca tcgtgttcaa ggccaagcgg atctacgact tcctgatccc cggcgcgccg
      781 tcgacctcca tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc
      841 aaccttggag aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct
      901 tcccagtcct ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact
      961 agcaacccca agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc
     1021 tcccttcagt ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat
     1081 caggaagtga acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt
     1141 gaggtgatgg gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag
     1201 gccagggtct ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg
     1261 caatgtctac tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc
     1321 aaagggttga attccggcgg cggctccgga ggcggtaccg gtggcggcga cctagcccca
     1381 cagatgcttc gagaactcca ggagactaat gcggcgctgc aagacgtgag agagctcttg
     1441 cgacagcagg tcaaggagat caccttcctg aagaatacgg tgatggaatg tgacgcctgc
     1501 ggtctagaca gaaacctgcc tccactagca cctctgggcc cacacccaga aacgctggtg
     1561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1681 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1921 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2161 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gggccctgac
     2341 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     2401 aagggccctc accaccacca tcaccactaa ttaattaagg atcccccgac ctcgacctct
     2461 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     2521 ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag atctctagct
     2581 agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct gccccttctt
     2641 cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac tgggccctgt
     2701 tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt agttgacatc
     2761 cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg agcagacttt
     2821 gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct gaagctctta
     2881 caccaatgct gggggacatg tacctcccag gggcccagga agactacggg aggctacacc
     2941 aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga gggcattagc
     3001 aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag
     3061 tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat
     3121 atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat
     3181 gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta
     3241 gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc
     3301 tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg
     3361 tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg
     3421 gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc
     3481 aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg
     3541 gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac
     3601 acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa
     3661 gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga
     3721 cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc
     3781 aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg
     3841 cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa
     3901 cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta
     3961 atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg
     4021 ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg
     4081 gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat gggtagcata
     4141 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcataggct
     4201 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4261 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4321 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     4381 atcctaatag agattagggt agtatatgct atcctaattt atatctgggt agcatatact
     4441 acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca tatgctatcc
     4501 taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     4561 taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc
     4621 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     4681 taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca tgagaatttt
     4741 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4801 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4861 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4921 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4981 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5041 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5101 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5161 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     5221 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5281 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5341 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5401 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5461 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     5521 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5581 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5641 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5701 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5761 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5821 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5881 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5941 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6121 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6541 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6601 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6661 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6721 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6781 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6841 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6901 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6961 gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca tgtttgacag
     7021 cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga actggtaggt
     7081 atggaagatc tatacattga atcaatattg gcaattagcc atattagtca ttggttatat
     7141 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     7201 catttatatt ggctcatgtc caatatgacc gccatgttga cattgattat tgactagtta
     7261 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     7321 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     7381 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     7441 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc
     7501 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     7561 cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     7621 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc
     7681 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7741 tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg
     7801 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcctcactc tcttccgcat
     7861 cgctgtctgc gagggccagc tgttgggctc gcggttgagg acaaactctt cgcggtcttt
     7921 ccagtactct tggatcggaa acccgtcggc ctccgaacgg tactccgcca ccgagggacc
     7981 tgagcgagtc cgcatcgacc ggatcggaaa acct
//
