LOCUS       Exported                8015 bp ds-DNA     circular SYN 13-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_0929900-COMP-blac-flag-his
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8015)
  TITLE     Pentamerised full-length protein ectodomains with no N-linked 
            glycans
REFERENCE   2  (bases 1 to 8015)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8015
                     /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             1549..2334
                     /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             2341..2406
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2413..2430
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2466..2521
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2520..2539)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4757..4775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4843..4947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4948..5808
                     /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      5979..6567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6468..6487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6721..6738
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6855..6876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6891..6921
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6929..6945
                     /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     6934..6956
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6953..6969
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6953..6969
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7240..7619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7620..7823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7774..7794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7820..7844
                     /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 ggccgcctac
      421 atgaacatca agagataccc caacaaggtg ctgaacagcg ccgagagcaa gttctacgac
      481 cacaacttcc acaagaacta catcctgaac aacctgttca agcggaacaa gctgtactac
      541 atctacccca agcagaacct gctgatcaag aacacccgga tctacaacgt gcccagagcc
      601 aagaccgcca tcagcctgtt taaggacatc aacatggcca actttttccc cggcttcaaa
      661 acaatcttca acaccaagtt cctcttcgac cctctgtaca gcagccacaa ggatatcatc
      721 atcaagaata tcaagaccat ccagctgctc cagcagggca actacgtgaa gtccggcgcg
      781 ccgtcgacct ccatcacggc ctataagagt gagggggagt cagcggagtt ctccttccca
      841 ctcaaccttg gagaggaaag cctgcaggga gagttgagat ggaaggcaga gaaggctcct
      901 tcttcccagt cctggatcac cttctcccta aagaaccaaa aggtgtctgt gcagaagtct
      961 actagcaacc ccaagttcca gctgtccgaa acgctcccac tcacccttca gataccccag
     1021 gtctcccttc agtttgctgg ttctggcaac ctgaccctga ctctggacag agggatactg
     1081 tatcaggaag tgaacctggt ggtgatgaaa gtgactcagc ccgacagcaa cactttgacc
     1141 tgtgaggtga tgggacccac ctcacccaag atgagactga tcttgaagca ggagaatcag
     1201 gaggccaggg tctccaggca ggagaaagtg attcaagtgc aggcccctga agcaggggtg
     1261 tggcaatgtc tactgagtga aggtgaagag gtcaagatgg actccaagat ccaggtttta
     1321 tccaaagggt tgaattccgg cggcggctcc ggaggcggta ccggtggcgg cgacctagcc
     1381 ccacagatgc ttcgagaact ccaggagact aatgcggcgc tgcaagacgt gagagagctc
     1441 ttgcgacagc aggtcaagga gatcaccttc ctgaagaata cggtgatgga atgtgacgcc
     1501 tgcggtctag acagaaacct gcctccacta gcacctctgg gcccacaccc agaaacgctg
     1561 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1621 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1681 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1741 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1801 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1861 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1921 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1981 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2041 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2101 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2161 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2221 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2281 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggggccct
     2341 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     2401 gacaagggcc ctcaccacca ccatcaccac taataataag gatcccccga cctcgacctc
     2461 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     2521 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc
     2581 tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct
     2641 tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg
     2701 ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat
     2761 ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt
     2821 tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt
     2881 acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac
     2941 caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag
     3001 caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta
     3061 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     3121 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca
     3181 tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt
     3241 agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc
     3301 ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag
     3361 gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac
     3421 ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg
     3481 caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg
     3541 ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa
     3601 cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca
     3661 agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg
     3721 acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc
     3781 caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg
     3841 gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata
     3901 acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact
     3961 aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt
     4021 gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt
     4081 ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat
     4141 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc
     4201 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4261 tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4321 tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc
     4381 tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac
     4441 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc
     4501 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4561 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     4621 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4681 ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt
     4741 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4801 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4861 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4921 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4981 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5041 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5101 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5161 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5221 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5281 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5341 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5401 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5461 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5521 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5581 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5641 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5701 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5761 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5821 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5881 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5941 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6001 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6061 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6121 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6181 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6241 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6301 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6361 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6421 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6481 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6541 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6601 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6661 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6721 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6781 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6841 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6901 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6961 agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca
     7021 gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg
     7081 tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     7141 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     7201 acatttatat tggctcatgt ccaatatgac cgccatgttg acattgatta ttgactagtt
     7261 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     7321 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     7381 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     7441 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc
     7501 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     7561 ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     7621 tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc
     7681 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     7741 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
     7801 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcctcact ctcttccgca
     7861 tcgctgtctg cgagggccag ctgttgggct cgcggttgag gacaaactct tcgcggtctt
     7921 tccagtactc ttggatcgga aacccgtcgg cctccgaacg gtactccgcc accgagggac
     7981 ctgagcgagt ccgcatcgac cggatcggaa aacct
//
