LOCUS       sequence-424371-        7984 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 7984)
  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 7984)
  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-FCGR1A-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1823..1858
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1868..1891
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3551..3595
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3596..3622
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3762..3779)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3768..3879
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3997..4017)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4002..4331
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4182..4317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4244..4263
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4379..4426
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4445..5044
                     /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(4445..4464)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4941..4961
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5174..5295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5211..5230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5265..5284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5348..6139,6140..6208))
                     /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     5971..5990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6209..6300)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6342..6361
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6367..6386
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6472..7060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6961..6980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7214..7231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7329..7346
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7330..7346
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7802..7818
                     /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)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg cccaagtgga tacaacaaag gccgtgatca ccctgcagcc
     1021 tccatgggtg tccgtgttcc aagaggaaac cgtgacactg cactgcgagg tgctgcatct
     1081 gcctggatct agcagcaccc agtggtttct gaatggcacc gccacacaga ccagcacacc
     1141 cagctataga atcaccagcg ccagcgtgaa cgacagcggc gagtatagat gccagagagg
     1201 cctgagcggc agaagcgacc ctattcagct ggaaatccac agaggctggc tgctgctgca
     1261 ggtttccagc agagtgttta cagagggcga gcccctggct ctgagatgtc acgcttggaa
     1321 ggacaagctg gtgtacaacg tgctgtacta ccggaacggc aaggccttca agttcttcca
     1381 ctggaacagc aacctgacca tcctcaagac caacatcagc cacaacggca cctaccactg
     1441 tagcggcatg ggcaagcaca gatacacaag cgccggcatc agcgtgaccg tgaaagagct
     1501 gtttcccgct cctgtgctga acgcctctgt gacaagtccc ctgctggaag gcaatctggt
     1561 caccctgtct tgcgagacaa agctgctgct tcagaggcct ggcctgcagc tgtacttcag
     1621 cttctacatg ggcagcaaga ccctgcgggg cagaaatacc agcagcgagt accagatcct
     1681 gaccgccaga agagaggaca gcggactgta ttggtgcgag gctgccaccg aggatggcaa
     1741 tgtgctgaag agaagccccg agctggaact gcaggttctg ggactgcaac tgcccacacc
     1801 tgtgtggttc catggcgcgc cggaagatca agttgatcca agattgattg atggcaaggg
     1861 cggatctctg gaggtgctgt tccagggacc aggcggaagc ggcaatcccg gcggcgatct
     1921 ggccccccag atgctgcgcg agctgcagga gaccaatgcc gccctgcagg acgtgcgcga
     1981 gctgctgcgc caccgcgtga aggagatcac cttcctgaag aacaccgtga tggagtgcga
     2041 cgcctgcggt atgcagccct ctagaggctc cggctccatc atcccagttg aggaggagaa
     2101 cccggacttc tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc
     2161 tgcacagaca gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac
     2221 ggtgacagct gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc
     2281 cctggccatg gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca
     2341 tgtgccagac agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca
     2401 gaccattggc ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga
     2461 ggtcatctcc gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac
     2521 cacacgagtg cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg
     2581 gtactcggac gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac
     2641 gcagctcatc tccaacatgg acattgacgt gatcctaggt ggaggccgaa agtacatgtt
     2701 tcgcatggga accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct
     2761 ggacgggaag aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg
     2821 gaaccgcact gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct
     2881 ctttgagcct ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct
     2941 gatggagatg acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct
     3001 cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact
     3061 gactgagacg atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga
     3121 ggacacgctg agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc
     3181 cctgcgaggg agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta
     3241 cacggtcctc ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga
     3301 tgttaccgag agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga
     3361 cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct
     3421 ggttcacggc gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct
     3481 ggagccctac accgcctgcg acctggcgcc ccccgccggc accaccgacg ccgcgcaccc
     3541 gggtggcgga ggactgaacg acatcttcga ggctcagaaa atcgaatggc acgaacatca
     3601 ccatcaccat caccatcacc attaaggccg gcctgtagtc actggcacaa cgcgttaccc
     3661 ggtaagccaa tcgggtatac acggtcgtca tactgcagac agggttcttc tactttgcaa
     3721 gatagtcttg agtagtaaaa taaatagata gagaaaatca gcctcgactg tgccttctag
     3781 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3841 tcccactgtc ctttcctaat aaaatgagga aattgcatca caacactcaa ccctatctcg
     3901 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3961 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4021 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4081 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4261 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4321 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     4381 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     4441 aaccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     4501 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     4561 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4621 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4681 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4741 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     4801 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     4861 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     4921 cctggagaca tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     4981 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5041 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5101 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5161 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5221 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5281 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5341 atggtcatta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5401 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5461 ctggccccag cgctgcgatg ataccgcgag aaccacgctc accggctccg gatttatcag
     5521 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5581 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5641 tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5701 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5761 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5821 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5881 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5941 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6001 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6061 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6121 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6181 gggcgacacg gaaatgttga atactcatat tcttcctttt tcaatattat tgaagcattt
     6241 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6301 taggggtcag tgttacaacc aattaaccaa ttctgaacat tatcgcgagc ccatttatac
     6361 ctgaatatgg ctcataacac cccttgctca tgaccaaaat cccttaacgt gagttacgcg
     6421 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6601 gcagatacca aatactgttc ttctagtgta gccgtagtta gcccaccact tcaagaactc
     6661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7081 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7141 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7201 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa ggcgagagta gggaactgcc
     7261 aggcatcaaa ctaagcagaa ggcccctgac ggatggcctt tttgcgtttc tacaaactct
     7321 ttctgtgttg taaaacgacg gccagtctta agctcgggcc ccctgggcgg ttctgataac
     7381 gagtaatcgt taatccgcaa ataacgtaaa aacccgcttc ggcgggtttt tttatggggg
     7441 gagtttaggg aaagagcatt tgtcagaata tttaagggcg cctgtcactt tgcttgatat
     7501 atgagaatta tttaacctta taaatgagaa aaaagcaacg cactttaaat aagatacgtt
     7561 gctttttcga ttgatgaaca cctataatta aactattcat ctattattta tgattttttg
     7621 tatatacaat atttctagtt tgttaaagag aattaagaaa ataaatctcg aaaataataa
     7681 agggaaaatc agtttttgat atcaaaatta tacatgtcaa cgataataca aaatataata
     7741 caaactataa gatgttatca gtatttatta tcatttagaa taaattttgt gtcgccctta
     7801 attgtgagcg gataacaatt acgagcttca tgcacagtgg cgttgactca atattggcca
     7861 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     7921 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     7981 tgtt
//
