LOCUS       sequence-385992-        8425 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 8425)
  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 8425)
  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-LILRA6-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8425
                     /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             2264..2299
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2309..2332
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3992..4036
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4037..4063
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(4203..4220)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4209..4320
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4438..4458)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4443..4772
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4623..4758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4685..4704
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4820..4867
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4886..5485
                     /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(4886..4905)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5382..5402
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5615..5736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5652..5671)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5706..5725
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5789..6580,6581..6649))
                     /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     6412..6431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6650..6741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6783..6802
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6808..6827
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6913..7501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7402..7421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7655..7672
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7770..7787
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7771..7787
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8243..8259
                     /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 ccggaccttt tcctaaacct acactgtggg ccgagcctgg
     1021 cagcgtgata tcttggggaa gccctgtgac catctggtgc cagggatctc tggaagccca
     1081 agagtaccag ctggacaaag agggctctcc cgagcctctg gacagaaaca acccactgga
     1141 acctaagaac aaggcccggt tcagcatccc cagcatgaca cagcatcacg ccggcagata
     1201 ccggtgccac tactatagtt ccgccggatg gtccgagcct agcgaccctc tggaactggt
     1261 catgacaggc ttctacaaca agcccacact gagcgccctg ccttctcctg ttgttgcctc
     1321 tggcggcaac atgaccctga gatgtggcag ccagaagggc taccaccact tcgtgctgat
     1381 gaaggaaggc gagcaccagc tgcctagaac actggattct cagcagctgc acagcggcgg
     1441 cttccaggct ctttttcctg tgggacctgt gacacccagc cacagatggc ggttcacctg
     1501 ttactactac tacaccaaca caccccgcgt gtggtcacac ccttccgatc ctctggaaat
     1561 cctgccttcc ggcgtgtcca gaaagcctag cctgctgaca ctgcagggac cagttctggc
     1621 tcctggccag tctctgactc tgcagtgtgg ctctgacgtg ggctacgaca gattcgtgct
     1681 gtacaaagaa ggcgagagag acttcctgca gaggcctgga cagcagcctc aagctggact
     1741 gagccaggcc aatttcacac tgggccctgt gtctccatct cacggcggcc agtacagatg
     1801 ttacggcgcc cacaatctga gcagcgagtg gtccgctcct agcgatcccc tgaacattct
     1861 gatggccgga cagatctacg acaccgtgtc tctgtctgcc cagcctggac ctacagtggc
     1921 cagcggagaa aatgtgaccc tgctgtgcca gagccggggc tacttcgata ccttcctgct
     1981 gaccaaagaa ggggccgctc atcctccact gcggctgaga tctatgtatg gggcccacaa
     2041 gtaccaggcc gagtttccta tgagccccgt gacatctgcc cacgcaggca cctatagatg
     2101 ctacggcagc tacagcagca accctcatct gctgagcttc cccagcgaac ccctcgagct
     2161 gatggtgtct ggacactctg gcggatctag cctgcctcct actggacctc caagcacacc
     2221 agcctctcac gccaaggact acaccgtgga aaatggcgcg ccggaagatc aagttgatcc
     2281 aagattgatt gatggcaagg gcggatctct ggaggtgctg ttccagggac caggcggaag
     2341 cggcaatccc ggcggcgatc tggcccccca gatgctgcgc gagctgcagg agaccaatgc
     2401 cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg aaggagatca ccttcctgaa
     2461 gaacaccgtg atggagtgcg acgcctgcgg tatgcagccc tctagaggct ccggctccat
     2521 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     2581 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     2641 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     2701 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     2761 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     2821 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     2881 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     2941 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     3001 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     3061 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     3121 tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     3181 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     3241 ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     3301 tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     3361 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     3421 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     3481 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     3541 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     3601 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     3661 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     3721 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     3781 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     3841 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     3901 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     3961 caccaccgac gccgcgcacc cgggtggcgg aggactgaac gacatcttcg aggctcagaa
     4021 aatcgaatgg cacgaacatc accatcacca tcaccatcac cattaaggcc ggcctgtagt
     4081 cactggcaca acgcgttacc cggtaagcca atcgggtata cacggtcgtc atactgcaga
     4141 cagggttctt ctactttgca agatagtctt gagtagtaaa ataaatagat agagaaaatc
     4201 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4261 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4321 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4381 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4441 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4501 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4561 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4621 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4681 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4741 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4801 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     4861 atacgacaag gtgaggaact aaaccatgac cgagtacaag cccacggtgc gcctcgccac
     4921 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4981 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5041 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5101 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5161 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5221 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5281 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5341 gcgcgccggg gtgcccgcct tcctggagac atccgcgccc cgcaacctcc ccttctacga
     5401 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5461 catgacccgc aagcccggtg cctgacacgt gctacgagat ttcgattcca ccgccgcctt
     5521 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5581 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5641 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5701 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5761 tagctagagc ttggcgtaat catggtcatt accaatgctt aatcagtgag gcacctatct
     5821 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5881 cgatacggga gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct
     5941 caccggctcc ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6001 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6061 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt
     6121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6421 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt
     6661 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6721 gtatttagaa aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca
     6781 ttatcgcgag cccatttata cctgaatatg gctcataaca ccccttgctc atgaccaaaa
     6841 tcccttaacg tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     6901 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6961 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7021 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7081 agcccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7141 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7201 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7261 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7321 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7381 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7441 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7501 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7561 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     7621 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     7681 aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga cggatggcct
     7741 ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc
     7801 cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa aaacccgctt
     7861 cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat atttaagggc
     7921 gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga aaaaagcaac
     7981 gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt aaactattca
     8041 tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga gaattaagaa
     8101 aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt atacatgtca
     8161 acgataatac aaaatataat acaaactata agatgttatc agtatttatt atcatttaga
     8221 ataaattttg tgtcgccctt aattgtgagc ggataacaat tacgagcttc atgcacagtg
     8281 gcgttgactc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     8341 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     8401 tcatgtccaa tatgaccgcc atgtt
//
