LOCUS       sequence-385993-        8473 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 8473)
  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 8473)
  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-LILRB2-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8473
                     /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             2312..2347
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2357..2380
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4040..4084
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4085..4111
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(4251..4268)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4257..4368
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4486..4506)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4491..4820
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4671..4806
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4733..4752
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4868..4915
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4934..5533
                     /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(4934..4953)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5430..5450
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5663..5784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5700..5719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5754..5773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5837..6628,6629..6697))
                     /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     6460..6479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6698..6789)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6831..6850
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6856..6875
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6961..7549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7450..7469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7703..7720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7818..7835
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7819..7835
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8291..8307
                     /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 cccaaactgg aacaatcccc aagcctacac tgtgggccga
     1021 gcctgatagc gtgatcacac agggaagccc cgtgacactg tcttgtcagg gatctctgga
     1081 agcccaagag taccggctgt acagagagaa gaagtccgcc agctggatca cccggatcag
     1141 acctgagctg gtcaagaacg gccagtttca catccccagc atcacctggg agcacaccgg
     1201 cagatacggc tgccagtact atagcagagc ccgttggagc gagctgagcg atcctctggt
     1261 gctggttatg accggcgcct atcctaagcc aacactgagc gcacagccct ctccagtggt
     1321 tacatctggc ggcagagtga ccctgcagtg cgaatctcag gtggccttcg gcggcttcat
     1381 cctgtgcaaa gagggcgaag aggaacaccc tcagtgcctg aatagccagc ctcacgccag
     1441 aggcagcagc agagccatct tttctgtggg ccccgtgtct cccaatcgga gatggtccca
     1501 cagatgctac ggctacgacc tgaacagccc ctacgtgtgg tctagcccta gcgacctgct
     1561 ggaactgctg gttcctggcg tgtccaagaa acccagcctg tctgttcagc ccggacctgt
     1621 tgttgcccct ggcgaatctc tgactctgca gtgtgtgtcc gacgtgggct acgacagatt
     1681 cgtgctgtac aaagaaggcg agcgggacct gagacagctg cctggtagac aacctcaggc
     1741 cggactgagc caggccaatt tcacactggg acccgtgtcc agaagctatg gcggccagta
     1801 cagatgttat ggcgcccaca atctgagcag cgagtgtagc gctcctagcg atcccctgga
     1861 catcctgatc accggccaga tcagaggcac ccctttcatc agtgtgcagc ctggacctac
     1921 agtggccagc ggcgaaaatg tgacactgct gtgccagtct tggcggcagt tccacacctt
     1981 tctgctgacc aaagccggcg ctgctgatgc ccctctgaga ctgagatcca tccacgagta
     2041 ccccaagtac caggccgagt ttcctatgag ccctgtgaca tctgcccacg ccggcaccta
     2101 tagatgttac ggcagcctga actctgaccc ctacctgctg tctcacccta gcgaaccact
     2161 cgagctggtg gtgtctggcc cttctatggg aagcagccca cctccaacag gccctatctc
     2221 tacaccagcc ggacctgagg atcagcccct gacaccaaca ggcagcgatc cacagtctgg
     2281 actgggcaga catctcggcg ttggcgcgcc ggaagatcaa gttgatccaa gattgattga
     2341 tggcaagggc ggatctctgg aggtgctgtt ccagggacca ggcggaagcg gcaatcccgg
     2401 cggcgatctg gccccccaga tgctgcgcga gctgcaggag accaatgccg ccctgcagga
     2461 cgtgcgcgag ctgctgcgcc accgcgtgaa ggagatcacc ttcctgaaga acaccgtgat
     2521 ggagtgcgac gcctgcggta tgcagccctc tagaggctcc ggctccatca tcccagttga
     2581 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     2641 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     2701 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     2761 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     2821 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     2881 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     2941 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     3001 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     3061 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     3121 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg gaggccgaaa
     3181 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     3241 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     3301 gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tgacccatct
     3361 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     3421 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     3481 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     3541 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     3601 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     3661 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     3721 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     3781 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     3841 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     3901 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     3961 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     4021 cgcgcacccg ggtggcggag gactgaacga catcttcgag gctcagaaaa tcgaatggca
     4081 cgaacatcac catcaccatc accatcacca ttaaggccgg cctgtagtca ctggcacaac
     4141 gcgttacccg gtaagccaat cgggtataca cggtcgtcat actgcagaca gggttcttct
     4201 actttgcaag atagtcttga gtagtaaaat aaatagatag agaaaatcag cctcgactgt
     4261 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4321 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcac aacactcaac
     4381 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4441 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4501 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4621 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4681 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4741 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4801 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4861 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4921 gaggaactaa accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     4981 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     5041 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5101 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5161 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     5221 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5281 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     5341 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     5401 gcccgccttc ctggagacat ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     5461 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     5521 gcccggtgcc tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     5581 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5641 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5701 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5761 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5821 ggcgtaatca tggtcattac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5941 gcttaccatc tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg
     6001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6121 ttaatagttt gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt
     6181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6421 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6661 agggaataag ggcgacacgg aaatgttgaa tactcatatt cttccttttt caatattatt
     6721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6781 ataaacaaat aggggtcagt gttacaacca attaaccaat tctgaacatt atcgcgagcc
     6841 catttatacc tgaatatggc tcataacacc ccttgctcat gaccaaaatc ccttaacgtg
     6901 agttacgcgc gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6961 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7021 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7081 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag cccaccactt
     7141 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7201 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7261 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7321 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7381 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7441 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7501 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7561 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7621 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     7681 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag gcgagagtag
     7741 ggaactgcca ggcatcaaac taagcagaag gcccctgacg gatggccttt ttgcgtttct
     7801 acaaactctt tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt
     7861 tctgataacg agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt
     7921 ttatgggggg agtttaggga aagagcattt gtcagaatat ttaagggcgc ctgtcacttt
     7981 gcttgatata tgagaattat ttaaccttat aaatgagaaa aaagcaacgc actttaaata
     8041 agatacgttg ctttttcgat tgatgaacac ctataattaa actattcatc tattatttat
     8101 gattttttgt atatacaata tttctagttt gttaaagaga attaagaaaa taaatctcga
     8161 aaataataaa gggaaaatca gtttttgata tcaaaattat acatgtcaac gataatacaa
     8221 aatataatac aaactataag atgttatcag tatttattat catttagaat aaattttgtg
     8281 tcgcccttaa ttgtgagcgg ataacaatta cgagcttcat gcacagtggc gttgactcaa
     8341 tattggccat tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg
     8401 ccattgcata cgttgtatct atatcataat atgtacattt atattggctc atgtccaata
     8461 tgaccgccat gtt
//
