LOCUS       sequence-405792-        8923 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 8923)
  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 8923)
  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-ILDR1-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8923
                     /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             1424..1459
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1469..1492
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3152..3196
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3197..3223
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3363..3380)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3369..3480
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3598..3618)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3603..3932
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3783..3918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3845..3864
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3980..4027
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4046..4645
                     /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(4046..4065)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4542..4562
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4775..4896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4812..4831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4866..4885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4949..5740,5741..5809))
                     /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     5572..5591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5810..5901)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5943..5962
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5968..5987
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6073..6661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6562..6581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6815..6832
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6930..6947
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6931..6947
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7403..7419
                     /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 ccttgctggt tacagtgcag cacaccgaga gatacgtgac
     1021 cctgttcgcc agcatcatcc tgaagtgcga ctacaccaca agcgcccagc tgcaggatgt
     1081 ggtggtcact tggagattca agagcttctg caaggacccc atcttcgact actacagcgc
     1141 cagctatcag gccgctctgt ctctgggcca agatcctagc aacgactgca acgacaacca
     1201 gcgggaagtg cggatcgttg ctcagagaag aggccagaat gagcctgtgc tgggcgtcga
     1261 ctaccggcag agaaagatca ccatccagaa ccgggccgac ctggtcatca acgaagtcat
     1321 gtggtgggac cacggcgtgt actactgtac aattgaggcc cctggcgaca ccagcggcga
     1381 tcctgacaaa gaagtgaagc tgatcgtcct gcatggcgcg ccggaagatc aagttgatcc
     1441 aagattgatt gatggcaagg gcggatctct ggaggtgctg ttccagggac caggcggaag
     1501 cggcaatccc ggcggcgatc tggcccccca gatgctgcgc gagctgcagg agaccaatgc
     1561 cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg aaggagatca ccttcctgaa
     1621 gaacaccgtg atggagtgcg acgcctgcgg tatgcagccc tctagaggct ccggctccat
     1681 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     1741 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     1801 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     1861 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     1921 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     1981 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     2041 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     2101 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     2161 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     2221 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     2281 tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     2341 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     2401 ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     2461 tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     2521 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     2581 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     2641 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     2701 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     2761 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     2821 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     2881 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     2941 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     3001 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     3061 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     3121 caccaccgac gccgcgcacc cgggtggcgg aggactgaac gacatcttcg aggctcagaa
     3181 aatcgaatgg cacgaacatc accatcacca tcaccatcac cattaaggcc ggcctgtagt
     3241 cactggcaca acgcgttacc cggtaagcca atcgggtata cacggtcgtc atactgcaga
     3301 cagggttctt ctactttgca agatagtctt gagtagtaaa ataaatagat agagaaaatc
     3361 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3421 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3481 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3541 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3601 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3661 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3721 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3781 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3841 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3901 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3961 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     4021 atacgacaag gtgaggaact aaaccatgac cgagtacaag cccacggtgc gcctcgccac
     4081 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4141 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4201 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4261 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4321 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4381 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4441 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4501 gcgcgccggg gtgcccgcct tcctggagac atccgcgccc cgcaacctcc ccttctacga
     4561 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4621 catgacccgc aagcccggtg cctgacacgt gctacgagat ttcgattcca ccgccgcctt
     4681 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4741 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4801 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4861 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4921 tagctagagc ttggcgtaat catggtcatt accaatgctt aatcagtgag gcacctatct
     4981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5041 cgatacggga gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct
     5101 caccggctcc ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt
     5281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt
     5821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5881 gtatttagaa aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca
     5941 ttatcgcgag cccatttata cctgaatatg gctcataaca ccccttgctc atgaccaaaa
     6001 tcccttaacg tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     6061 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6121 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6181 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6241 agcccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6301 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6361 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6421 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6481 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6541 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6601 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6661 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6721 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6781 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6841 aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga cggatggcct
     6901 ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc
     6961 cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa aaacccgctt
     7021 cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat atttaagggc
     7081 gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga aaaaagcaac
     7141 gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt aaactattca
     7201 tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga gaattaagaa
     7261 aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt atacatgtca
     7321 acgataatac aaaatataat acaaactata agatgttatc agtatttatt atcatttaga
     7381 ataaattttg tgtcgccctt aattgtgagc ggataacaat tacgagcttc atgcacagtg
     7441 gcgttgactc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     7501 attggctatt ggccattgca tacgttgtac tgagagatcc cctcataatt tccccaaaac
     7561 gtaaccatgt gtgaatagat tttgagtaag cagggttgca gccacgagtg agtcttccct
     7621 tgttattgtg tagccagaat gccgcaaaac ttccatgcct aagcgaactg ttgagagtac
     7681 gtttcgattt ctgactgtgt tagcctggaa gtgcttgtcc caaccttgtt tctgagcatg
     7741 aacgcccgca agccaacatg ttagttgaag catcagggcg attagcagca tgatatcaaa
     7801 acgctctgag ctgctcgttc ggctatggcg taggcctagt ccgtaggcag gacttttcaa
     7861 gtctcggaag gtttcttcaa tctgcattcg cttcgaatag atattaacaa gttgtttggg
     7921 tgttcgaatt tcaacaggta agttagttgc tagaatccat ggctcctttg ccgacgctga
     7981 gtagatttta ggtgacgggt ggtgacaatg agtccgtgtc gagcgctgat tttttcggcc
     8041 tttagagcga gatttataca atagaatttg gcatgagatt gggttgcttt tagtcagcct
     8101 cttatagcct aaagtctttg agtgactaga tgacatatca tgtaagttgc tgataggttt
     8161 ccagttttcc gctcctaggt ctgcatattg tacttttcct cttactcgac ttaaccagta
     8221 ccaacccagc ttctcaacgg atttatacca tggcacttta aagccagcat cactgacaat
     8281 gagcggtgtg gtgttactcg gtagaatgct cgcaaggtcg gctagaaatt ggtcatgagc
     8341 tttctttgaa cattgctctg aaagcgggaa cgctttctca taaagagtaa cagaacgacc
     8401 gtgtagtgcg actgaagctc gcaataccat aagccgtttt tgctcacgga tatcagacca
     8461 gtcaacaagt acaatgggca tcgtattgcc cgaacagata aagctagcat gccaacggta
     8521 tacagcgagt cgctctttgt ggaggtgacg attacctaac aatcggtcga ttcgtttgat
     8581 gttatgtttt gttctcgctt tggttggcag gttacggcca agttcggtaa gagtgagagt
     8641 tttacagtca agtaaggcgt ggcaagccaa cgttaagctg ttgagtcgtt ttaagtgtaa
     8701 ttcggggcag aattggtaaa gagagtcgtg taaaatatcg agttcgcaca ttttgttgtc
     8761 tgattattga tttttggcga aaccatttga tcatatgaca agatgtgtat ctaccttaac
     8821 ttaatgattt tgataaaaat cattagggga ttcatcagta cgttgtatct atatcataat
     8881 atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
