LOCUS       Exported                8971 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    PSOP24-COMP-blac-flag-his
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8971)
  TITLE     Pentamerised full-length protein ectodomains with no N-linked 
            glycans
REFERENCE   2  (bases 1 to 8971)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8971
                     /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             2503..3288
                     /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             3295..3360
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3367..3384
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3422..3477
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3476..3495)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5713..5731)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5799..5903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5904..6764
                     /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      6935..7523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7424..7443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7677..7694
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7811..7832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7847..7877
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7885..7901
                     /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     7890..7912
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7909..7925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7909..7925
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8196..8575
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8576..8779
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8730..8750
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8776..8800
                     /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 ggccgccaag
      421 aacatcaacc tgaacgacga gggcaagaag aagcagagcg gcatcatcaa cgagaatgag
      481 gacggcaaga acaacaaggc caacaacaac aagaaggtgc agcacaacaa gatcaaccgg
      541 aagggcaaca acgccgtgac caagaaaacc gacgagcaca agaacgacgg cgacgccgag
      601 aacaagaaga atggcgagca caaaaatgat ggggacggga aaaacaagaa aaacggggaa
      661 cataagaacg atggggatgg aaagaacaaa aagatggatg agcacctgta cgacatcgac
      721 ggggagaata agaaaaatgg tgaacacctc tacgatgtgg acggcgagaa caaaaaaaac
      781 ggcgaacacc tgtatgatat cgatggcgaa aacaaaaaga acggggagca tctctacgac
      841 attgatggcg agaacaagaa aacagatgag cacctgtatg acgtggacgg cggcaaaaac
      901 aacatcctcg agtacaacaa catcgaccag cacaacgagt tccccgagag cctggaaaac
      961 gacaacagct acgacaagct gttcgacgac gtggaactgc gggacatcat ccacaacgag
     1021 aagttcttcg agaacctgaa gaacgtgaac aacaacgacg tgcacaactt tctgctggtg
     1081 gacaaagaga tggaacgccg caaagaggaa gagaagaaga agtcccggaa gctggaagag
     1141 gacgaagagg atgacgagga cgaggaagaa gataacgacg agaagtggaa agaggaaaat
     1201 aagaacaacg ctgccagcag cagctcctcc agcagcaaca acttcaacaa gaactacgac
     1261 atctacgaca accgggacct gtacaactac gataacctgg tggatatcac ccaggaagaa
     1321 cacatcaacg tccccaacat caacgatgag atcaacgagg gccggaagca cgacaaagag
     1381 gactttctga tctttgagaa gcggctgaac gaggccgagg accggaacga cgccagcgac
     1441 ggggataaca agatggaaga tctgatcaac aaccacttcg acgacagcgg cgacaacggc
     1501 gcctacgaga acatccaggt caaaaaagag ctgtacaaca aaaacgagaa gatcatcaag
     1561 ggccgggaca acaacctgaa tgatggcgac atccacatga acaaaaacaa tgccgccaac
     1621 aaactcaaca acaataacaa caacaacaac aatgacaaca atatgaacag cgagatgaag
     1681 tacaagacct tcaaagaaga gtacgaggaa cagatcctga gcaagcccgg cgcgccgtcg
     1741 acctccatca cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac
     1801 cttggagagg aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc
     1861 cagtcctgga tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc
     1921 aaccccaagt tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc
     1981 cttcagtttg ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag
     2041 gaagtgaacc tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag
     2101 gtgatgggac ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc
     2161 agggtctcca ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa
     2221 tgtctactga gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa
     2281 gggttgaatt ccggcggcgg ctccggaggc ggtaccggtg gcggcgacct agccccacag
     2341 atgcttcgag aactccagga gactaatgcg gcgctgcaag acgtgagaga gctcttgcga
     2401 cagcaggtca aggagatcac cttcctgaag aatacggtga tggaatgtga cgcctgcggt
     2461 ctagacagaa acctgcctcc actagcacct ctgggcccac acccagaaac gctggtgaaa
     2521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattgggg ccctgactac
     3301 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     3361 ggccctcacc accaccatca ccactaatta attaaggatc ccccgacctc gacctctggc
     3421 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3481 ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc tctagctaga
     3541 ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc ccttcttcgc
     3601 ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg gccctgttcc
     3661 acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt tgacatcctt
     3721 ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc agactttgca
     3781 gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac
     3841 caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac
     3901 gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat
     3961 agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc cgggtagtag
     4021 tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg
     4081 ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc accccatgag
     4141 ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac cattttagtc
     4201 acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc
     4261 ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac agtaaggtgt
     4321 atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat ttggacgggg
     4381 ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc cttgggcaat
     4441 aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat ccatggggtg
     4501 gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat gggcaacaca
     4561 taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag ggaccaagac
     4621 aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg acgccgacag
     4681 cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct ccacgccaat
     4741 ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga gtgggggcac
     4801 gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt gtaataactt
     4861 ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa accactaatg
     4921 gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc gcgattgctg
     4981 cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg gttgttggtc
     5041 ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg tagcatatac
     5101 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc ataggctatc
     5161 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     5221 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     5281 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     5341 ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc atatactacc
     5401 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat gctatcctaa
     5461 tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     5521 tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa
     5581 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     5641 tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga gaattttctt
     5701 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5761 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5821 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5881 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5941 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6001 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6061 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6121 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     6181 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6241 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6301 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6361 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6421 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     6481 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6541 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6601 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6661 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6721 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6781 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6841 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6901 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6961 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7021 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7081 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7141 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7201 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7261 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7321 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7381 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7441 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7501 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7561 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7621 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7681 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     7741 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     7801 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     7861 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     7921 atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt ttgacagctt
     7981 atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact ggtaggtatg
     8041 gaagatctat acattgaatc aatattggca attagccata ttagtcattg gttatatagc
     8101 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
     8161 ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga ctagttatta
     8221 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     8281 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     8341 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8401 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     8461 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     8521 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     8581 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     8641 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     8701 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     8761 ggtctatata agcagagctc gtttagtgaa ccgtcagatc ctcactctct tccgcatcgc
     8821 tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca aactcttcgc ggtctttcca
     8881 gtactcttgg atcggaaacc cgtcggcctc cgaacggtac tccgccaccg agggacctga
     8941 gcgagtccgc atcgaccgga tcggaaaacc t
//
