LOCUS       Exported                8017 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 8017)
  TITLE     Pentamerised full-length protein ectodomains with no N-linked 
            glycans
REFERENCE   2  (bases 1 to 8017)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8017
                     /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    2468..2523
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2522..2541)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4759..4777)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4845..4949
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4950..5810
                     /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      5981..6569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6470..6489
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6723..6740
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6857..6878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6893..6923
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6931..6947
                     /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     6936..6958
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6955..6971
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6955..6971
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7242..7621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7622..7825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7776..7796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7822..7846
                     /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 taattaatta aggatccccc gacctcgacc
     2461 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     2521 tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta
     2581 gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt
     2641 cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc
     2701 tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac
     2761 atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac
     2821 tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc
     2881 ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac
     2941 accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt
     3001 agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg
     3061 tagtagtata tactatccag actaacccta attcaatagc atatgttacc caacgggaag
     3121 catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc
     3181 catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt
     3241 ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc
     3301 gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta
     3361 aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg
     3421 acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg
     3481 ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat
     3541 ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc
     3601 aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac
     3661 caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc
     3721 cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac
     3781 gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg
     3841 gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa
     3901 taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca
     3961 ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga
     4021 ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg
     4081 ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc
     4141 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag
     4201 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4261 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4321 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     4381 gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat
     4441 actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta
     4501 tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     4561 tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta
     4621 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     4681 tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat
     4741 tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4801 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4921 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4981 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5041 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5101 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5161 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     5221 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5281 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5341 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5401 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5461 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     5521 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5581 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5641 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5701 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5761 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5821 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5881 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6121 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6601 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6661 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6721 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     6781 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     6841 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     6901 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     6961 acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc tcatgtttga
     7021 cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta
     7081 ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta
     7141 tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat
     7201 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
     7261 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     7321 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     7381 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     7441 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     7501 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     7561 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     7621 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
     7681 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     7741 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
     7801 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctca ctctcttccg
     7861 catcgctgtc tgcgagggcc agctgttggg ctcgcggttg aggacaaact cttcgcggtc
     7921 tttccagtac tcttggatcg gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg
     7981 acctgagcga gtccgcatcg accggatcgg aaaacct
//
