LOCUS       Exported                7809 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses full-length Adipocyte plasma membrane-associated protein 
            ectodomain in mammalian cells. N-terminal His tag, biotinylation 
            sequence and rat Cd4d3+4 tag..
ACCESSION   .
VERSION     .
KEYWORDS    C20-His-bio
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7809)
  AUTHORS   Sun Y, Vandenbriele C, Kauskot A, Verhamme P, Hoylaerts MF, Wright 
            GJ
  TITLE     A human platelet receptor protein microarray identifies 
            FcepsilonR1alpha as an activating PEAR1 ligand.
  JOURNAL   Mol Cell Proteomics. 2015 Feb 23. pii: mcp.M114.046946.
  PUBMED    25713122
REFERENCE   2  (bases 1 to 7809)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7809
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      1..10
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             70..87
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1909..1964
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1963..1982)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4200..4218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4286..4390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4391..5251
                     /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"
     primer_bind     complement(4609..4628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5422..6010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5911..5930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6164..6181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6298..6319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6334..6364
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6372..6388
                     /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     6377..6399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6396..6412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6396..6412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6683..7062
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7063..7266
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7217..7237
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7263..7287
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     7768..7787
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gccaccatgg agtttcagac ccaggtactc atgtccctgc tgctctgcat gtctggtctg
       61 aattccggcc accaccatca ccaccatagc cccgacagga acctgcctcc tctggcccct
      121 ctgggcccca acagcggcag cccagacctg catcatattc tggatgcaca gaaaatggtg
      181 tggaatcatc gtgacagaaa cctgcctcca ctagcacctc tgtcgacctc catcacggcc
      241 tataagagtg agggggagtc agcggagttc tccttcccac tcaaccttgg agaggaaagc
      301 ctgcagggag agttgagatg gaaggcagag aaggctcctt cttcccagtc ctggatcacc
      361 ttctccctaa agaaccaaaa ggtgtctgtg cagaagtcta ctagcaaccc caagttccag
      421 ctgtccgaaa cgctcccact cacccttcag ataccccagg tctcccttca gtttgctggt
      481 tctggcaacc tgaccctgac tctggacaga gggatactgt atcaggaagt gaacctggtg
      541 gtgatgaaag tgactcagcc cgacagcaac actttgacct gtgaggtgat gggacccacc
      601 tcacccaaga tgagactgat cttgaagcag gagaatcagg aggccagggt ctccaggcag
      661 gagaaagtga ttcaagtgca ggcccctgaa gcaggggtgt ggcaatgtct actgagtgaa
      721 ggtgaagagg tcaagatgga ctccaagatc caggttttat ccaaagggtt gaattcgcgg
      781 ccgcccattg atccccagcc cctgagcttc aaagagcccc ccctgctgct gggagtgctg
      841 caccccaaca ccaagctgcg gcaggccgag agactgttcg agaaccagct cgtgggaccc
      901 gagtctatcg cccacatcgg cgacgtgatg ttcaccggca ccgccgacgg cagagtggtc
      961 aagctggaaa acggcgagat cgagacaatc gccagattcg gcagcggccc ctgcaagacc
     1021 agagatgacg agcccgtgtg cggcagacct ctgggcatca gagccggccc taacggcacc
     1081 ctgttcgtgg ccgacgccta caagggcctg ttcgaagtga acccctggaa gcgggaagtg
     1141 aagctgctcc tgtccagcga gacacccatc gagggcaaga acatgagctt cgtgaacgac
     1201 ctgacagtga cccaggacgg ccggaagatc tacttcaccg atagcagcag caagtggcag
     1261 cggcgggact atctgctgct ggtcatggaa ggcaccgacg atggccggct gctggaatac
     1321 gacaccgtga cccgggaggt caaggtgctg ctggaccagc tgagattccc caacggcgtg
     1381 cagctgtccc ctgccgagga tttcgtgctg gtcgccgaga caaccatggc cagaatccgg
     1441 cgggtgtacg tgtctggcct gatgaagggc ggagccgacc tgtttgtgga gaacatgccc
     1501 ggcttccccg acaacatcag acctagcagc tctggcggct actgggtcgg catgagcacc
     1561 atcagaccca accccggctt cagcatgctg gactttctga gcgagcggcc ctggatcaag
     1621 cggatgatct tcaagctgtt cagccaagag acagtgatga agttcgtgcc ccggtacagc
     1681 ctggtgctgg aactgagcga cagcggcgcc ttcagaagaa gcctgcacga ccccgatggc
     1741 ctggtggcta cctacatcag cgaggtgcac gagcacgacg gccacctgta cctgggcagc
     1801 ttccggtccc ccttcctgtg cagactgtct ctgcaggccg tgtgaggcgc gcctgaattc
     1861 ttctagacac gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat
     1921 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     1981 ggagggcaaa tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg
     2041 ccccggacga actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg
     2101 taatcccttc agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg
     2161 ggggggacca aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg
     2221 cacatttgcc aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc
     2281 aacacaacat tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga
     2341 catgtacctc ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg
     2401 gcctgtgtag ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg
     2461 cccccttgtt aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca
     2521 gactaaccct aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg
     2581 gttagtaaaa gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt
     2641 atttacatgg ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg
     2701 ctgaagatca aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc
     2761 ttcgtttagc taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc
     2821 gaaaacaagg tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg
     2881 cattgtgcta tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta
     2941 ggaatgaaac attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta
     3001 aagactggat gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca
     3061 atatgatact ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt
     3121 tgttacactc tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac
     3181 tggttgtctc taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac
     3241 aaagacaagt ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca
     3301 cacgccgccc tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac
     3361 cccgctaacc actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa
     3421 tacctgcata agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac
     3481 aaattacaca cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga
     3541 ggtcgctgag agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct
     3601 ggatagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     3661 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     3721 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     3781 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta
     3841 gggtagtata tgctatccta atttatatct gggtagcata tactacccaa atatctggat
     3901 agcatatgct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3961 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4021 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4081 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4141 agcatatgct atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg
     4201 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4381 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4501 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4561 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4621 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4681 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4801 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4921 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4981 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5221 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5281 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5341 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6061 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6181 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6241 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6301 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6361 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6421 cgccaagctc tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc
     6481 cgggcaacgt tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca
     6541 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
     6601 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     6661 tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
     6721 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6781 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6841 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6901 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
     6961 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
     7021 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     7081 tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     7141 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat
     7201 aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     7261 agagctcgtt tagtgaaccg tcagatcctc actctcttcc gcatcgctgt ctgcgagggc
     7321 cagctgttgg gctcgcggtt gaggacaaac tcttcgcggt ctttccagta ctcttggatc
     7381 ggaaacccgt cggcctccga acggtactcc gccaccgagg gacctgagcg agtccgcatc
     7441 gaccggatcg gaaaacctct cgagaaaggc gtctaaccag tcacagtcgc aaggtaggct
     7501 gagcaccgtg gcgggcggca gcgggtggcg gtcggggttg tttctggcgg aggtgctgct
     7561 gatgatgtaa ttaaagtagg cggtcttgag acggcggatg gtcgaggtga ggtgtggcag
     7621 gcttgagatc cagctgttgg ggtgagtact ccctctcaaa agcgggcatt acttctgcgc
     7681 taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc gatctggcca
     7741 tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg
     7801 tccaagttt
//
