LOCUS       Exported                8680 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_0910300-COMP-blac-flag-his
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8680)
  TITLE     Pentamerised full-length protein ectodomains with no N-linked 
            glycans
REFERENCE   2  (bases 1 to 8680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8680
                     /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             2212..2997
                     /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             3004..3069
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3076..3093
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3131..3186
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3185..3204)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5422..5440)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5508..5612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5613..6473
                     /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      6644..7232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7133..7152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7386..7403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7520..7541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7556..7586
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7594..7610
                     /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     7599..7621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7618..7634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7618..7634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7905..8284
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8285..8488
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8439..8459
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8485..8509
                     /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 ggccgccgag
      421 gaattcgaga agatcaagta ccctcaggac gtgaccaaga ttaagtacta cgacaacagc
      481 cacaagatcg acagcaagta cctgtacaag cacgtggcca agggcatgct gcaggacctg
      541 atcgtgaccg tgcagatcga caaggacaga gaggtggaca agatcctgtt cctgcccaag
      601 aaaaagacct acatgaagat catcggcgat tggagcgact accccaaaga gaagaacaac
      661 gagatctgcc tggccttcgt gctgcagaag tactttaaga aagagaaaga aaaaaagtgc
      721 accaaggatt tcttcaccag cggcaacatc tacaacgtgt gctacaacga caacaactac
      781 gccatcaaga acctgtacct ctacatcgag atcagccaga agtacctcga gaagctgttc
      841 agcagctaca gcatccccgt gatcgtgaac accgagaaca acatcatcta cttcgccaac
      901 gacaagatga agtccaaaga ggaaaagccc atcctggacc tgcacatctg ctacaagagc
      961 aactacgaga cacccaccta cctgggcaag ctgatcttca actcctaccc catcaacatc
     1021 aaccggaaca aggaccagtt cgacattatc tacaccaaga gctttctgac caacaactgg
     1081 gactacaacc tggccatcaa gggcaactac atcaccagct acaaccgggt ggtgttcgtg
     1141 cggatcccca gagacagccc tcccaacaaa atgaagtgcc cacccttctg ggattacagc
     1201 gtgctgggca gcggccctct gcacaagatc aagaagaaaa acaagaacaa tatcattcag
     1261 aacatggtgc agtacagctt cagcaacaag aatatccaga actactacat cccattcctc
     1321 aagaaggacg agctgcagac caacgaggcc tacatcatct gcctgtacag cgacgagaac
     1381 gacttcgagg gcatgcagat catcaacgcc cacttctaca tcaacaccgc cgacagcggc
     1441 gcgccgtcga cctccatcac ggcctataag agtgaggggg agtcagcgga gttctccttc
     1501 ccactcaacc ttggagagga aagcctgcag ggagagttga gatggaaggc agagaaggct
     1561 ccttcttccc agtcctggat caccttctcc ctaaagaacc aaaaggtgtc tgtgcagaag
     1621 tctactagca accccaagtt ccagctgtcc gaaacgctcc cactcaccct tcagataccc
     1681 caggtctccc ttcagtttgc tggttctggc aacctgaccc tgactctgga cagagggata
     1741 ctgtatcagg aagtgaacct ggtggtgatg aaagtgactc agcccgacag caacactttg
     1801 acctgtgagg tgatgggacc cacctcaccc aagatgagac tgatcttgaa gcaggagaat
     1861 caggaggcca gggtctccag gcaggagaaa gtgattcaag tgcaggcccc tgaagcaggg
     1921 gtgtggcaat gtctactgag tgaaggtgaa gaggtcaaga tggactccaa gatccaggtt
     1981 ttatccaaag ggttgaattc cggcggcggc tccggaggcg gtaccggtgg cggcgaccta
     2041 gccccacaga tgcttcgaga actccaggag actaatgcgg cgctgcaaga cgtgagagag
     2101 ctcttgcgac agcaggtcaa ggagatcacc ttcctgaaga atacggtgat ggaatgtgac
     2161 gcctgcggtc tagacagaaa cctgcctcca ctagcacctc tgggcccaca cccagaaacg
     2221 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2281 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2341 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2401 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2461 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2521 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2581 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2641 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2701 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2761 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2821 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2881 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2941 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggggc
     3001 cctgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
     3061 gatgacaagg gccctcacca ccaccatcac cactaattaa ttaaggatcc cccgacctcg
     3121 acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     3181 ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc tcggagatct
     3241 ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac atctctgccc
     3301 cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt gccaactggg
     3361 ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct gactgtagtt
     3421 gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca tcctggagca
     3481 gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc ttggctgaag
     3541 ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac tacgggaggc
     3601 tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc tcaagagggc
     3661 attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca tatgcttccc
     3721 gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt acccaacggg
     3781 aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg atatctccca
     3841 ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg gtagtgaacc
     3901 attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact ctcctgaatc
     3961 ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga gttgtgaaca
     4021 gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt
     4081 tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct cacaaacccc
     4141 ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac aatagaaatc
     4201 catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt tatggctatg
     4261 ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt cccaggcagg
     4321 gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa agagagtgga
     4381 cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta aacggggctc
     4441 cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga aattgtggag
     4501 tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa aataagggtg
     4561 taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt gcccacaaaa
     4621 ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa gataggggcg
     4681 cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc aagcacaggg
     4741 ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt tgccatgggt
     4801 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     4861 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4921 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4981 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     5041 tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat ctgggtagca
     5101 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatatg
     5161 ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     5221 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg
     5281 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     5341 ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt caaacatgag
     5401 aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5461 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5521 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5581 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5641 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5701 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5761 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5821 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     5881 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5941 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6001 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6061 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6121 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     6181 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6241 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6301 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6361 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6421 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6481 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6541 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6601 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6661 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6721 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6781 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6841 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6901 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6961 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7021 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7081 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7141 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7201 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7261 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7321 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7381 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7441 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7501 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     7561 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     7621 gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt
     7681 tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg cgcagaactg
     7741 gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat tagtcattgg
     7801 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
     7861 tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac
     7921 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     7981 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     8041 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     8101 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     8161 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     8221 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     8281 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
     8341 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     8401 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
     8461 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcc tcactctctt
     8521 ccgcatcgct gtctgcgagg gccagctgtt gggctcgcgg ttgaggacaa actcttcgcg
     8581 gtctttccag tactcttgga tcggaaaccc gtcggcctcc gaacggtact ccgccaccga
     8641 gggacctgag cgagtccgca tcgaccggat cggaaaacct
//
