LOCUS       Exported                8134 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    CelTOS-COMP-blac-flag-his
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8134)
  TITLE     Pentamerised full-length protein ectodomains with no N-linked 
            glycans
REFERENCE   2  (bases 1 to 8134)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8134
                     /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             1666..2451
                     /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             2458..2523
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2530..2547
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2585..2640
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2639..2658)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4876..4894)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4962..5066
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5067..5927
                     /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      6098..6686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6587..6606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6840..6857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6974..6995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7010..7040
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7048..7064
                     /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     7053..7075
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7072..7088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7072..7088
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7359..7738
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7739..7942
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7893..7913
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7939..7963
                     /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 ggccgccttc
      421 agaggcaaca acggccacaa cagcgccagc agcctgtaca acggcgccca gttcatcgag
      481 cagctgaaca acgccttcac cagcgccttt ctggaaagcc agagcatgaa caagatcggc
      541 gacgacctgg ccgagacaat cagcaacgag ctggtgtccg tgctgcagaa gaacagcccc
      601 accttcctgg aatccagctt cgacatcaag agcgaagtga agaaacacgc caagagcatg
      661 ctgaaagagc tgatcaaagt cggcctgcct agcttcgaga acctggtggc cgagaacgtg
      721 aagcctccca aggtggaccc cgccacctac ggcatcatcg tgcccgtgct gaccagcctg
      781 ttcaacaagg tggaaaccgc cgtgggcgcc aaggtgtccg acgagatctg gaactacaac
      841 agccccgacg tgtccgagag cgaggaaagc ctgagcgacg acttcttcga cggcgcgccg
      901 tcgacctcca tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc
      961 aaccttggag aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct
     1021 tcccagtcct ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact
     1081 agcaacccca agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc
     1141 tcccttcagt ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat
     1201 caggaagtga acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt
     1261 gaggtgatgg gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag
     1321 gccagggtct ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg
     1381 caatgtctac tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc
     1441 aaagggttga attccggcgg cggctccgga ggcggtaccg gtggcggcga cctagcccca
     1501 cagatgcttc gagaactcca ggagactaat gcggcgctgc aagacgtgag agagctcttg
     1561 cgacagcagg tcaaggagat caccttcctg aagaatacgg tgatggaatg tgacgcctgc
     1621 ggtctagaca gaaacctgcc tccactagca cctctgggcc cacacccaga aacgctggtg
     1681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gggccctgac
     2461 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     2521 aagggccctc accaccacca tcaccactaa ttaattaagg atcccccgac ctcgacctct
     2581 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     2641 ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag atctctagct
     2701 agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct gccccttctt
     2761 cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac tgggccctgt
     2821 tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt agttgacatc
     2881 cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg agcagacttt
     2941 gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct gaagctctta
     3001 caccaatgct gggggacatg tacctcccag gggcccagga agactacggg aggctacacc
     3061 aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga gggcattagc
     3121 aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag
     3181 tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat
     3241 atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat
     3301 gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta
     3361 gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc
     3421 tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg
     3481 tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg
     3541 gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc
     3601 aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg
     3661 gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac
     3721 acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa
     3781 gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga
     3841 cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc
     3901 aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg
     3961 cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa
     4021 cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta
     4081 atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg
     4141 ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg
     4201 gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat gggtagcata
     4261 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcataggct
     4321 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4381 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4441 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     4501 atcctaatag agattagggt agtatatgct atcctaattt atatctgggt agcatatact
     4561 acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca tatgctatcc
     4621 taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     4681 taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc
     4741 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     4801 taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca tgagaatttt
     4861 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4921 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4981 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5041 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5101 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5161 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5221 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5281 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     5341 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5401 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5461 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5521 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5581 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     5641 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5701 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5761 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5821 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5881 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5941 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6001 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6061 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6121 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6181 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6241 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6301 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6361 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6421 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6481 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6541 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6601 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6661 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6721 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6781 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6841 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6901 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6961 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     7021 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     7081 gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca tgtttgacag
     7141 cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga actggtaggt
     7201 atggaagatc tatacattga atcaatattg gcaattagcc atattagtca ttggttatat
     7261 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     7321 catttatatt ggctcatgtc caatatgacc gccatgttga cattgattat tgactagtta
     7381 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     7441 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     7501 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     7561 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc
     7621 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     7681 cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     7741 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc
     7801 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7861 tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg
     7921 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcctcactc tcttccgcat
     7981 cgctgtctgc gagggccagc tgttgggctc gcggttgagg acaaactctt cgcggtcttt
     8041 ccagtactct tggatcggaa acccgtcggc ctccgaacgg tactccgcca ccgagggacc
     8101 tgagcgagtc cgcatcgacc ggatcggaaa acct
//
