LOCUS       sequence-420996-        8005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to COMP5AP-AviTag-9xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8005)
  AUTHORS   Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL, 
            Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser 
            SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC
  TITLE     A Human IgSF Cell-Surface Interactome Reveals a Complex Network of 
            Protein-Protein Interactions.
  JOURNAL   Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 
            10.1016/j.cell.2020.07.025.
  PUBMED    32822567
REFERENCE   2  (bases 1 to 8005)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2020.07"; journalName: "Cell"; date: "2020-08-20- 
            20"; volume: "182"; issue: "4"; pages: "1027-1043.e17"
COMMENT     Sequence Label: pD649-HAsp-MICA-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             313..357
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             358..384
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(524..541)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    530..641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(759..779)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        764..1093
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      944..1079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1006..1025
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1141..1188
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1207..1806
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1207..1226)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1703..1723
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1936..2057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1973..1992)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2027..2046
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(2110..2901,2902..2970))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2733..2752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2971..3062)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3104..3123
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3129..3148
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3234..3822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3723..3742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3976..3993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4091..4108
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4092..4108
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4564..4580
                     /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)."
     enhancer        4747..5126
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5127..5330
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5281..5301
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5327..5351
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        5561..5579
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5561..5578
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     5592..5611
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6590..6625
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             6635..6658
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg
       61 gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc agtgcccctg
      121 gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac
      181 ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt cgccgcctgc
      241 ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga cgccgcgcac
      301 ccgggtggcg gaggactgaa cgacatcttc gaggctcaga aaatcgaatg gcacgaacat
      361 caccatcacc atcaccatca ccattaaggc cggcctgtag tcactggcac aacgcgttac
      421 ccggtaagcc aatcgggtat acacggtcgt catactgcag acagggttct tctactttgc
      481 aagatagtct tgagtagtaa aataaataga tagagaaaat cagcctcgac tgtgccttct
      541 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
      601 actcccactg tcctttccta ataaaatgag gaaattgcat cacaacactc aaccctatct
      661 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      721 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
      781 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      841 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
      901 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
      961 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1021 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     1081 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     1141 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     1201 taaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     1261 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     1321 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     1381 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     1441 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     1501 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     1561 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     1621 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     1681 ttcctggaga catccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     1741 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     1801 gcctgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     1861 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     1921 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1981 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2041 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     2101 tcatggtcat taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2161 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2221 atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc cggatttatc
     2281 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2341 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2401 tttgcgcaac gttgttgcca tcgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2461 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2521 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2581 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2641 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2701 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2761 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2821 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2881 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2941 aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt attgaagcat
     3001 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3061 aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga gcccatttat
     3121 acctgaatat ggctcataac accccttgct catgaccaaa atcccttaac gtgagttacg
     3181 cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3361 gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca cttcaagaac
     3421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3901 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3961 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaggcgagag tagggaactg
     4021 ccaggcatca aactaagcag aaggcccctg acggatggcc tttttgcgtt tctacaaact
     4081 ctttctgtgt tgtaaaacga cggccagtct taagctcggg ccccctgggc ggttctgata
     4141 acgagtaatc gttaatccgc aaataacgta aaaacccgct tcggcgggtt tttttatggg
     4201 gggagtttag ggaaagagca tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat
     4261 atatgagaat tatttaacct tataaatgag aaaaaagcaa cgcactttaa ataagatacg
     4321 ttgctttttc gattgatgaa cacctataat taaactattc atctattatt tatgattttt
     4381 tgtatataca atatttctag tttgttaaag agaattaaga aaataaatct cgaaaataat
     4441 aaagggaaaa tcagtttttg atatcaaaat tatacatgtc aacgataata caaaatataa
     4501 tacaaactat aagatgttat cagtatttat tatcatttag aataaatttt gtgtcgccct
     4561 taattgtgag cggataacaa ttacgagctt catgcacagt ggcgttgact caatattggc
     4621 cattagccat attattcatt ggttatatag cataaatcaa tattggctat tggccattgc
     4681 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     4741 catgttggca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     4801 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4861 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4921 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4981 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     5041 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     5101 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     5161 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     5221 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     5281 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     5341 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
     5401 accgatccag cctccgcggc cgggaacggt gcattggaac gcggattccc cgtgccaaga
     5461 gtgacgtaag taccgcctat agagtctata ggcccacccc cttggcttcg ttagaacgcg
     5521 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa
     5581 taacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac tgcacctcgg
     5641 ttctatcgaa aacgcgcctg ctagcgccac catgaagacc atcattgccc tgagctacat
     5701 cttctgcctg gtgttcgctg cggccgccga acctcacagc ctgagataca atctgaccgt
     5761 gctgagctgg gatggcagcg tgcagtctgg atttctgacc gaagtgcacc tggacggcca
     5821 gcctttcctg agatgcgaca gacagaagtg cagagccaag cctcaaggcc agtgggctga
     5881 agatgtgctg ggcaacaaga cctgggacag agagacaaga gatctgaccg gcaacggcaa
     5941 ggacctgaga atgacactgg cccacatcaa ggaccagaaa gagggcctgc acagcctgca
     6001 agagatcaga gtgtgcgaga tccacgagga caacagcacc agaagcagcc agcacttcta
     6061 ctacgacggc gagctgttcc tgagccagaa cctggaaaca aaagaatgga ccatgcctca
     6121 gagcagcaga gcccagacac tggctatgaa cgtgcggaac ttcctgaaag aggacgccat
     6181 gaagaccaag acacactacc acgccatgca cgccgactgt ctgcaagagc tgagaagata
     6241 cctgaagtcc ggcgtggtgc tgagaaggac agtgcctcct atggtcaacg tgaccagatc
     6301 tgaggccagc gagggcaata tcaccgtgac atgtagagcc agcggcttct acccctggaa
     6361 catcaccctg tcttggagac aggatggcgt gtccctgagc cacgatacac agcagtgggg
     6421 agatgtcctg cctgacggca atggcacata ccagacatgg gtcgccacca gaatctgcca
     6481 gggcgaagaa cagcggttca cctgttacat ggaacacagc ggcaaccaca gcacacaccc
     6541 tgtgccatct ggaaaggtgc tggtgctgca gtctcactgg ggcgcgccgg aagatcaagt
     6601 tgatccaaga ttgattgatg gcaagggcgg atctctggag gtgctgttcc agggaccagg
     6661 cggaagcggc aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac
     6721 caatgccgcc ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt
     6781 cctgaagaac accgtgatgg agtgcgacgc ctgcggtatg cagccctcta gaggctccgg
     6841 ctccatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     6901 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     6961 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     7021 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     7081 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     7141 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     7201 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     7261 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     7321 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     7381 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     7441 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     7501 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     7561 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     7621 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     7681 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     7741 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     7801 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     7861 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     7921 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     7981 tggcaaggcc cgggacagga aggcc
//
