LOCUS       Exported                9677 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    DPAP1-COMP-blac-flag-his
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9677)
  TITLE     Pentamerised full-length protein ectodomains with no N-linked 
            glycans
REFERENCE   2  (bases 1 to 9677)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9677
                     /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             3211..3996
                     /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             4003..4068
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4075..4092
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4128..4183
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4182..4201)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6419..6437)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6505..6609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6610..7470
                     /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      7641..8229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8130..8149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8383..8400
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8517..8538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8553..8583
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8591..8607
                     /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     8596..8618
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8615..8631
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8615..8631
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8902..9281
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9282..9485
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9436..9456
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9482..9506
                     /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 ggccgccgat
      421 ctgcccaccc acgtggaaac aaagaacctg ctgggcaagt ggaagatcct gcggaccaag
      481 accagcccca acctggccac ctgtggcagc agccagccca acaagaacac ctacaacgtg
      541 ggcatcaccg actacaagaa gtacctgctg gaaaacaact acgagttcgt gtccgagctg
      601 aacgtgatcc tgagcgacga ctacgtgctg tacggcgaca tctacaacac ccaggacaac
      661 gagcaccggt ctaagtggaa ggtgctggcc gtgtacgacg agaacaagag agtgatcggc
      721 acctggacca ccatctgcga cgagggcttc gagatcaaga tcggcaacga ggcctacgcc
      781 gccctgatgc actacgagcc caacggcaag tgcggccctg tgtccgacga ggacagcctg
      841 gacagcaacg gcgacaccga ctgctacacc accagcttca gcaagatcag atacggctgg
      901 ctggacgtgg aaaacgagaa gaacgagcat ctgcacggct gcttctacgc cgagcggatc
      961 ttcgacaacg tgaacgagat taagcacctc gacagcttca ccatcgacaa ggactcccag
     1021 aacgtgctgc agaccttcac ctacgacacc aagctgaaca acatcctgaa cagcaacaac
     1081 atgctgtaca agttcggcaa cctgcagaag cccaccttca ccaagagaaa caacgccaac
     1141 gtgcagttca acagcgagct gaattggcac cggatgaagc accacggcaa gaagaagccc
     1201 ctgaaaaaga gcatgctgga cgccagccgg cagacctacg cctgcccctg caacgccaat
     1261 gaggtggtgg acaatgtgat caacaagggc gacagcgaca accccgtgtc ccctaccctg
     1321 atccagctca acaacaatct gaagaacacc gcccagaccg gcaacaagga caccaacgag
     1381 atggacctgg aaaattacga ggacacactg aacagcccca agcgcgagct ggaaatcaac
     1441 gagctgccca agaacttcgc ctggggcgac ccctggaaca agaatacccg cgagtacgaa
     1501 gtgaccaacc agctgctgtg cggcagctgc tatatcgcca gccagctgta cgccttcaag
     1561 cggcggatcg aggtggccct gaccaagaag ctggacagaa agtacctgaa caacttcgac
     1621 gaccagctga gcatccagac cgtgctgtcc tgcagcttct acgaccaggg ctgcaacggc
     1681 ggcttcccct acctggtgtc caagctggcc aaactgcagg gcatccctct gaacgtgtac
     1741 ttcccttaca gcgccaccga ggaaacctgc ccctacaata tcgccaagca ccccaacgac
     1801 atgaacggcg ctgccaagct gagagagatc aacgccatct tcaactctaa caacaatatg
     1861 gccacataca acaacatcaa caacgaccat caccagctgg gagtgtacgc caacaccgcc
     1921 agctcccagg aacagcacgg catcagcgaa gagaaccggt ggtacgccaa ggacttcaac
     1981 tacgtgggcg gctgctacgg ctgcaaccag tgcaatggcg agaagatcat gatgaatgag
     2041 atctaccgga acggccccat cgtgtctagc ttcgaggcca gccccgactt ctacgattac
     2101 gccgacggcg tgtactttgt ggaagatttc cctcacgcca gacggtgcac catcgagccc
     2161 aagaatgatg gcgtgtacaa cattgccgga tgggacagag tgaaccacgc catcgtgctg
     2221 ctgggctggg gcgaggaaga gattaacggc aagctgtata agtactggat cggccggaac
     2281 agctggggca acggctgggg aaaagagggc tacttcaaga tcctgagagg ccagaatttc
     2341 gccggcatcg agagccagag cctgttcatc gagcctgact tcagccgggg agccggcaag
     2401 attctgctcg agaagatgca gaaagagctg ggaaacggcg cgccgtcgac ctccatcacg
     2461 gcctataaga gtgaggggga gtcagcggag ttctccttcc cactcaacct tggagaggaa
     2521 agcctgcagg gagagttgag atggaaggca gagaaggctc cttcttccca gtcctggatc
     2581 accttctccc taaagaacca aaaggtgtct gtgcagaagt ctactagcaa ccccaagttc
     2641 cagctgtccg aaacgctccc actcaccctt cagatacccc aggtctccct tcagtttgct
     2701 ggttctggca acctgaccct gactctggac agagggatac tgtatcagga agtgaacctg
     2761 gtggtgatga aagtgactca gcccgacagc aacactttga cctgtgaggt gatgggaccc
     2821 acctcaccca agatgagact gatcttgaag caggagaatc aggaggccag ggtctccagg
     2881 caggagaaag tgattcaagt gcaggcccct gaagcagggg tgtggcaatg tctactgagt
     2941 gaaggtgaag aggtcaagat ggactccaag atccaggttt tatccaaagg gttgaattcc
     3001 ggcggcggct ccggaggcgg taccggtggc ggcgacctag ccccacagat gcttcgagaa
     3061 ctccaggaga ctaatgcggc gctgcaagac gtgagagagc tcttgcgaca gcaggtcaag
     3121 gagatcacct tcctgaagaa tacggtgatg gaatgtgacg cctgcggtct agacagaaac
     3181 ctgcctccac tagcacctct gggcccacac ccagaaacgc tggtgaaagt aaaagatgct
     3241 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3301 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3361 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3421 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3481 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3541 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3601 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3661 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3721 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3781 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3841 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3901 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3961 atcgctgaga taggtgcctc actgattaag cattggggcc ctgactacaa agaccatgac
     4021 ggtgattata aagatcatga catcgattac aaggatgacg atgacaaggg ccctcaccac
     4081 caccatcacc actaataata aggatccccc gacctcgacc tctggctaat aaaggaaatt
     4141 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     4201 gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc
     4261 cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt
     4321 aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg
     4381 gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc
     4441 acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca
     4501 acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac
     4561 atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca atcagagggg
     4621 cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc
     4681 ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag
     4741 actaacccta attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg
     4801 ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta
     4861 tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc
     4921 tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct
     4981 tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg
     5041 aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc
     5101 attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag
     5161 gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa
     5221 agactggatg tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa
     5281 tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt
     5341 gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact
     5401 ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca
     5461 aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac
     5521 acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc
     5581 ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat
     5641 acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca
     5701 aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag
     5761 gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg
     5821 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     5881 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     5941 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     6001 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag
     6061 ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata
     6121 gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     6181 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     6241 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     6301 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     6361 gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc
     6421 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6481 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6541 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6601 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6661 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6721 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6781 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6841 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6901 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6961 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7021 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7081 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7141 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7201 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7261 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7321 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7381 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7441 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7501 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7561 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7621 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7681 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7741 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7801 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7861 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7921 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7981 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8041 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8101 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8161 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8221 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8281 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8341 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8401 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8461 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8521 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8581 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8641 gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     8701 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     8761 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     8821 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     8881 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
     8941 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     9001 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     9061 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     9121 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
     9181 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
     9241 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     9301 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     9361 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
     9421 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     9481 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc
     9541 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg
     9601 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcga gtccgcatcg
     9661 accggatcgg aaaacct
//
