LOCUS       sequence-421003-        8230 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 8230)
  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 8230)
  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-AMIGO2-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1115..1159
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1160..1186
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(1326..1343)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1332..1443
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1561..1581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1566..1895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1746..1881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1808..1827
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1943..1990
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2009..2608
                     /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(2009..2028)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2505..2525
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2738..2859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2775..2794)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2829..2848
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(2912..3703,3704..3772))
                     /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     3535..3554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3773..3864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3906..3925
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3931..3950
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4036..4624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4525..4544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4778..4795
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4893..4910
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4894..4910
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5366..5382
                     /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        5549..5928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5929..6132
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6083..6103
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6129..6153
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        6363..6381
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6363..6380
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6394..6413
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             7617..7652
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             7662..7685
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
       61 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
      121 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
      181 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
      241 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
      301 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
      361 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
      421 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
      481 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
      541 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
      601 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
      661 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
      721 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
      781 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
      841 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
      901 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
      961 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     1021 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     1081 cggcaccacc gacgccgcgc acccgggtgg cggaggactg aacgacatct tcgaggctca
     1141 gaaaatcgaa tggcacgaac atcaccatca ccatcaccat caccattaag gccggcctgt
     1201 agtcactggc acaacgcgtt acccggtaag ccaatcgggt atacacggtc gtcatactgc
     1261 agacagggtt cttctacttt gcaagatagt cttgagtagt aaaataaata gatagagaaa
     1321 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     1381 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     1441 atcacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     1561 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1621 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     1681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     1741 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     1801 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     1861 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     1921 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     1981 ataatacgac aaggtgagga actaaaccat gaccgagtac aagcccacgg tgcgcctcgc
     2041 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     2101 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     2161 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     2221 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     2281 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     2341 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     2401 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     2461 cgagcgcgcc ggggtgcccg ccttcctgga gacatccgcg ccccgcaacc tccccttcta
     2521 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     2581 gtgcatgacc cgcaagcccg gtgcctgaca cgtgctacga gatttcgatt ccaccgccgc
     2641 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     2701 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     2761 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2821 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     2881 ctctagctag agcttggcgt aatcatggtc attaccaatg cttaatcagt gaggcaccta
     2941 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3001 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     3061 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3121 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3181 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     3241 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3301 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3361 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3421 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3481 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3541 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3601 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3661 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3721 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     3781 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3841 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     3901 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg ctcatgacca
     3961 aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa
     4021 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4081 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4141 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4201 gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4261 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4321 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4381 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4441 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4501 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4561 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4621 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4681 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4741 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4801 ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc tgacggatgg
     4861 cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt cttaagctcg
     4921 ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg taaaaacccg
     4981 cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag aatatttaag
     5041 ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg agaaaaaagc
     5101 aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata attaaactat
     5161 tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa agagaattaa
     5221 gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa attatacatg
     5281 tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt attatcattt
     5341 agaataaatt ttgtgtcgcc cttaattgtg agcggataac aattacgagc ttcatgcaca
     5401 gtggcgttga ctcaatattg gccattagcc atattattca ttggttatat agcataaatc
     5461 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     5521 ggctcatgtc caatatgacc gccatgttgg cattgattat tgactagtta ttaatagtaa
     5581 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     5641 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     5701 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     5761 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     5821 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     5881 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     5941 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     6001 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     6061 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     6121 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     6181 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
     6241 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagagtcta taggcccacc
     6301 cccttggctt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata
     6361 cgatttaggt gacactatag aataacatcc actttgcctt tctctccaca ggtgtccact
     6421 cccaggtcca actgcacctc ggttctatcg aaaacgcgcc tgctagcgcc accatgaaga
     6481 ccatcattgc cctgagctac atcttctgcc tggtgttcgc tgcggccgcc gtttgtccaa
     6541 cagcctgtat ctgtgccacc gacatcgtgt cctgcaccaa caagaatctg agcaaggtgc
     6601 ccggcaacct gttccggctg atcaagagac tggacctgag ctacaaccgg atcggcctgc
     6661 tggatagcga gtggattcct gtgtccttcg ccaagctgaa caccctgatc ctgcggcaca
     6721 acaacatcac cagcatcagc accggcagct tcagcaccac acctaacctg aagtgcctgg
     6781 acctgtccag caacaagctg aaaaccgtga agaacgccgt gttccaagag ctgaaggtgc
     6841 tggaagtgct gctgctgtac aacaaccaca tcagctatct ggaccctagc gcctttggcg
     6901 gactgtctca gctgcagaag ctgtacctga gcggcaactt cctgacacag ttccccatgg
     6961 acctgtacgt gggcagattc aagctggccg agctgatgtt cctggacgtg tcctacaaca
     7021 gaatccccag catgcccatg caccacatca acctggttcc aggcaagcag ctgaggggca
     7081 tctatctgca cggcaaccct ttcgtgtgcg actgctctct gtacagcctg ctggtgttct
     7141 ggtacagacg gcacttctcc agcgtgatgg acttcaagaa cgactacacc tgtcggcttt
     7201 ggagcgacag cagacacagt agacaggtcc tgctgctgca agacagcttc atgaactgca
     7261 gcgactccat catcaacgga agcttcagag ccctgggctt catccacgag gcccaagtgg
     7321 gagaaagact gatggtgcac tgcgacagca agaccggcaa cgccaacacc gatttcatct
     7381 gggtcggacc cgacaacaga ctgctggaac ccgacaaaga gatggaaaac ttctacgtgt
     7441 tccacaacgg cagcctggtc atcgagagcc ctagatttga agatgccggc gtgtactcct
     7501 gtatcgccat gaacaagcag cggctgctga acgagacagt ggacgtgacc atcaacgtgt
     7561 ccaacttcac cgtgtccaga agccacgctc acgaggcctt caacacaggc gcgccggaag
     7621 atcaagttga tccaagattg attgatggca agggcggatc tctggaggtg ctgttccagg
     7681 gaccaggcgg aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc
     7741 aggagaccaa tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga
     7801 tcaccttcct gaagaacacc gtgatggagt gcgacgcctg cggtatgcag ccctctagag
     7861 gctccggctc catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag
     7921 ccgaggccct gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca
     7981 tcatcttcct gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag
     8041 ggcagaagaa ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg
     8101 tggctctgtc caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca
     8161 cggcctacct gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc
     8221 gctttaacca
//
