LOCUS       sequence-457773-        8428 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 8428)
  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 8428)
  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-SIGLEC5-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     465..484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     718..735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     833..850
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     834..850
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1306..1322
                     /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        1489..1868
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1869..2072
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2023..2043
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2069..2093
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2303..2321
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2303..2320
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2334..2353
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3755..3790
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             3800..3823
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5483..5527
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5528..5554
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(5694..5711)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5700..5811
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5929..5949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5934..6263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6114..6249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6176..6195
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6311..6358
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6377..6976
                     /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(6377..6396)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6873..6893
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7106..7227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7143..7162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7197..7216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7280..8071,8072..8140))
                     /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     7903..7922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8141..8232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8274..8293
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8299..8318
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(8404..8428,1..564)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
       61 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      121 gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc accgcctaca
      181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
      721 gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc
      781 agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac
      841 gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc
      901 gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt agggaaagag
      961 catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac
     1021 cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg
     1081 aacacctata attaaactat tcatctatta tttatgattt tttgtatata caatatttct
     1141 agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt
     1201 tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt
     1261 atcagtattt attatcattt agaataaatt ttgtgtcgcc cttaattgtg agcggataac
     1321 aattacgagc ttcatgcaca gtggcgttga ctcaatattg gccattagcc atattattca
     1381 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     1441 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat
     1501 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     1561 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     1621 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     1681 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     1741 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1801 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     1861 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     1921 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     1981 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     2041 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     2101 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg
     2161 gccgggaacg gtgcattgga acgcggattc cccgtgccaa gagtgacgta agtaccgcct
     2221 atagagtcta taggcccacc cccttggctt cgttagaacg cggctacaat taatacataa
     2281 ccttatgtat catacacata cgatttaggt gacactatag aataacatcc actttgcctt
     2341 tctctccaca ggtgtccact cccaggtcca actgcacctc ggttctatcg aaaacgcgcc
     2401 tgctagcgcc accatgaaga ccatcattgc cctgagctac atcttctgcc tggtgttcgc
     2461 tgcggccgcc gaaaagcctg tgtatgagct gcaggtccag aaaagcgtga ccgtgcaaga
     2521 gggcctgtgt gtgctggtgc cttgcagctt tagctaccct tggcggagct ggtacagcag
     2581 ccctcctctg tacgtgtact ggttcagaga tggcgagatc ccctactacg ccgaggtggt
     2641 ggccacaaac aaccccgaca gaagagtgaa gcccgagaca cagggcagat tcagactgct
     2701 gggcgacgtg cagaagaaga actgcagcct gagcatcggc gacgcccgga tggaagatac
     2761 cggcagctac ttcttcagag tggaacgcgg cagggacgtg aagtacagct accagcagaa
     2821 caagctgaac ctggaagtga cagccctgat cgagaagccc gacatccact ttctggaacc
     2881 cctggaaagc ggcagaccca ccagactgtc ttgtagcctg cctggcagct gtgaagctgg
     2941 ccctccactg acctttagct ggaccggcaa tgccctgtct cctctggacc ctgagacaac
     3001 cagaagcagc gagctgaccc tgacacctag acctgaggac cacggcacca atctgacctg
     3061 ccagatgaag aggcagggcg cccaagtgac caccgagaga acagtgcagc tgaacgtgtc
     3121 ctacgctccc cagaccatca ccatcttccg gaacggaatc gccctggaaa tcctgcagaa
     3181 caccagctac ctgcctgtgc tggaaggcca ggctctgaga ctgctgtgtg acgcccctag
     3241 caatcctcct gctcacctga gctggtttca gggcagccca gctctgaacg ccacacctat
     3301 cagcaacacc ggcatcctgg aactgagaag agtgcggagc gctgaggaag gcggcttcac
     3361 ttgtagagcc cagcatcctc tgggctttct gcagatcttc ctgaacctga gcgtgtacag
     3421 cctgccacag ctgctgggac ctagctgttc ttgggaagcc gagggcctgc actgccggtg
     3481 tagctttaga gctagacccg ctcctagcct gtgttggcgg ctggaagaga aacctctgga
     3541 aggcaatagc agccagggca gcttcaaagt gaacagcagc tctgccggac cttgggccaa
     3601 tagctctctg attctgcacg gcggcctgag cagcgatctg aaggtgtcat gcaaggcctg
     3661 gaacatctac ggcagccagt ctggctctgt gctgctgctt cagggaagaa gcaatctcgg
     3721 cacaggcgtg gtgcctgctg ctcttggcgc gccggaagat caagttgatc caagattgat
     3781 tgatggcaag ggcggatctc tggaggtgct gttccaggga ccaggcggaa gcggcaatcc
     3841 cggcggcgat ctggcccccc agatgctgcg cgagctgcag gagaccaatg ccgccctgca
     3901 ggacgtgcgc gagctgctgc gccaccgcgt gaaggagatc accttcctga agaacaccgt
     3961 gatggagtgc gacgcctgcg gtatgcagcc ctctagaggc tccggctcca tcatcccagt
     4021 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     4081 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     4141 gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg acaaactggg
     4201 gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca agacatacaa
     4261 tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt gcggggtcaa
     4321 gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt gcaacacgac
     4381 acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga agtcagtggg
     4441 agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg cccacacggt
     4501 gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg aggggtgcca
     4561 ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag gtggaggccg
     4621 aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact acagccaagg
     4681 tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc gccagggtgc
     4741 ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt ctgtgaccca
     4801 tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag actccacact
     4861 ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca ggaacccccg
     4921 cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg aaagcagggc
     4981 ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg cgggccagct
     5041 caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg tcttctcctt
     5101 cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca aggcccggga
     5161 caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg
     5221 cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc
     5281 agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc
     5341 gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt
     5401 cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga
     5461 cgccgcgcac ccgggtggcg gaggactgaa cgacatcttc gaggctcaga aaatcgaatg
     5521 gcacgaacat caccatcacc atcaccatca ccattaaggc cggcctgtag tcactggcac
     5581 aacgcgttac ccggtaagcc aatcgggtat acacggtcgt catactgcag acagggttct
     5641 tctactttgc aagatagtct tgagtagtaa aataaataga tagagaaaat cagcctcgac
     5701 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5761 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cacaacactc
     5821 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5941 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6061 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6181 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6241 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     6301 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     6361 ggtgaggaac taaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     6421 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     6481 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     6541 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     6601 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     6661 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     6721 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     6781 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     6841 ggtgcccgcc ttcctggaga catccgcgcc ccgcaacctc cccttctacg agcggctcgg
     6901 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     6961 caagcccggt gcctgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     7021 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7081 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7201 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7261 cttggcgtaa tcatggtcat taccaatgct taatcagtga ggcacctatc tcagcgatct
     7321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7381 agggcttacc atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc
     7441 cggatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7561 cagttaatag tttgcgcaac gttgttgcca tcgctacagg catcgtggtg tcacgctcgt
     7621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt
     8161 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8221 aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga
     8281 gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa atcccttaac
     8341 gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8401 ttcttgagat cctttttttc tgcgcgta
//
