LOCUS       sequence-420988-        7777 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 7777)
  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 7777)
  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-CD80-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(35..51)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(150..167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(321..909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(401..420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(995..1014)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1020..1039)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        1081..1172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1173..1241,1242..2033)
                     /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     complement(1391..1410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2086..2207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2097..2116)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2151..2170
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2337..2936)
                     /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(2420..2440)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2917..2936
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2955..3002)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3050..3379)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3064..3199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3118..3137)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3364..3384
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3502..3613
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3602..3619
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3759..3785)
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             complement(3786..3830)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(5490..5513)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(5523..5558)
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     primer_bind     complement(6309..6328)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(6341..6359)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6342..6359)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6569..6593)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6590..6793)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6619..6639)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6794..7173
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    7340..7356
                     /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 gttatcagaa ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa
       61 agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct
      121 ggcagttccc tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      721 gagttcttga agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc
      781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg
      961 cgcgtaactc acgttaaggg attttggtca tgagcaaggg gtgttatgag ccatattcag
     1021 gtataaatgg gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta
     1081 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1141 aaatgcttca ataatattga aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc
     1201 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1261 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1321 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1381 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1441 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1501 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1561 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1621 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1681 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc gatggcaaca acgttgcgca
     1741 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1801 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1861 ctgataaatc cggagccggt gagcgtggtt ctcgcggtat catcgcagcg ctggggccag
     1921 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1981 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taatgaccat
     2041 gattacgcca agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg
     2101 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2161 atgctattgc tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag
     2221 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt
     2281 ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtag cacgtgtcag
     2341 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     2401 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga tgtctccagg
     2461 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     2521 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     2581 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
     2641 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     2701 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     2761 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     2821 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     2881 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt
     2941 tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga ttaattgtca
     3001 acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc
     3061 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct
     3121 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     3181 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     3241 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     3301 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     3361 cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag
     3421 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3481 cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag gacagtggga
     3541 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
     3601 ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact caagactatc
     3661 ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga ttggcttacc
     3721 gggtaacgcg ttgtgccagt gactacaggc cggccttaat ggtgatggtg atggtgatgg
     3781 tgatgttcgt gccattcgat tttctgagcc tcgaagatgt cgttcagtcc tccgccaccc
     3841 gggtgcgcgg cgtcggtggt gccggcgggg ggcgccaggt cgcaggcggt gtagggctcc
     3901 aggcaggcgg cgaaggccat gacgtgcgct atgaaggtct gctcctgcac gccgtgaacc
     3961 aggtgcgcct gcgggccgcg cgcgaacacc gccacgtcct cgcctgcgtg ggtctcttcg
     4021 tccaggggca ctgctgactg ctgccgatac tcggggctcc cgctctcgct ctcggtaaca
     4081 tccggccggg cgccgtcctt gagcacatag cctggaccgt ttccgtatag gaggaccgtg
     4141 taggccttcc tgtcccgggc cttgccaggg gccagcccga agatggagct ccctcgcagg
     4201 gggtagcctc cgaaggagaa gacgtgggag tggtcggcag tgacgaggct cagcgtgtcc
     4261 tcctcgctgg tgagctggcc cgccctctca atggcgtcgt cgaacatgat cgtctcagtc
     4321 agtgcccggt aagccctgct ttcatgatga ccatggtcga tgcgaccacc ctccacgaag
     4381 aggaagaagc cgcgggggtt cctgctcagc aggcgcaggg cagcctctgt catctccatc
     4441 agggaggggt ccagtgtgga gtctcggtgg atctcgtatt tcatgtctcc aggctcaaag
     4501 agacccatga gatgggtcac agacgggtcc agggaagcct gcatgagctc agtgcggttc
     4561 cacacatacc gggcaccctg gcgcttcgcc agccattcct gcaccagatt cttcccgtcc
     4621 agcctggtcc caccttggct gtagtcatct gggtactcag ggtctggggt tcccatgcga
     4681 aacatgtact ttcggcctcc acctaggatc acgtcaatgt ccatgttgga gatgagctgc
     4741 gtagcgatgt cctggcaccc ctcctggcgg gccgaggcag gcacgtcggc gtccgagtac
     4801 cagttgcggt tcaccgtgtg ggcgtaggtg ccggctggcg aggcgtgctg cactcgtgtg
     4861 gtggttacca ctcccactga cttccctgct ttcttggccc gattcatcac ggagatgacc
     4921 tcgttgccgc gtgtcgtgtt gcactggtta aagcgggcgg ctgcactcaa gccaatggtc
     4981 tggaagttgc ccttgacccc gcacaggtag gccgtggctg tggctccact gtctggcaca
     5041 tgtttgtcta cattgtatgt cttggacaga gccacatatg ggaagcggtc catggccagg
     5101 ggtatctcag gccccagttt gtccttcttc tgccctttta ggatcctggc agctgtcacc
     5161 gtagacaccc ccatcccatc gcccaggaag atgatgaggt tcttggcggc tgtctgtgca
     5221 ggctgcagct tcttggcggc acccagggcc tcggctgcct cgcggttcca gaagtccggg
     5281 ttctcctcct caactgggat gatggagccg gagcctctag agggctgcat accgcaggcg
     5341 tcgcactcca tcacggtgtt cttcaggaag gtgatctcct tcacgcggtg gcgcagcagc
     5401 tcgcgcacgt cctgcagggc ggcattggtc tcctgcagct cgcgcagcat ctggggggcc
     5461 agatcgccgc cgggattgcc gcttccgcct ggtccctgga acagcacctc cagagatccg
     5521 cccttgccat caatcaatct tggatcaact tgatcttccg gcgcgccatt gtcggggaag
     5581 tgctcttgct tggtggtatt ccagttgaag gtctggttca ctctcaggtg gccgtactta
     5641 atcaggcaca tgaagctgtg gttggtggtc atgttgaagt ccagcttgct ggacacggca
     5701 tacagctctg tctcggggtc ctgagacacg gtggtgttga tggcgttcag ttcctcgccg
     5761 ttttccagcc aagacagatg aggctctgga aagccgccgc tggtgctaca gatgatccgt
     5821 ctgatgttgc tggtggggat ctcgaagtcg ctgatgctag gtgtgggaaa gtcggccttc
     5881 acgctcagtg tcacttcggc caggtgttcc cgcttgaagg cgtccttctc gtacttcagc
     5941 accacgcact cataggtgcc ctcatcagaa ggcctcaggg ccagaatcac gatgctcagg
     6001 ttgttggtga tgtcgaagat ggtccggttc ttgtactcgg gccagatgtt catgtcgccg
     6061 ctcatcatgg tcagcaccat tttcttttct ttctgccagt agatcctggt ctgggccagt
     6121 tcttccactg aaacgttgtg gccgcagctc agtgtggcga cctctttcac ttctttggtc
     6181 acgtggatca cggcggccgc agcgaacacc aggcagaaga tgtagctcag ggcaatgatg
     6241 gtcttcatgg tggcgctagc aggcgcgttt tcgatagaac cgaggtgcag ttggacctgg
     6301 gagtggacac ctgtggagag aaaggcaaag tggatgttat tctatagtgt cacctaaatc
     6361 gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac gaagccaagg
     6421 gggtgggcct atagactcta taggcggtac ttacgtcact cttggcacgg ggaatccgcg
     6481 ttccaatgca ccgttcccgg ccgcggaggc tggatcggtc ccggtgtctt ctatggaggt
     6541 caaaacagcg tggatggcgt ctccaggcga tctgacggtt cactaaacga gctctgctta
     6601 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     6661 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     6721 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     6781 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     6841 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     6901 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     6961 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     7021 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     7081 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     7141 attactatta ataactagtc aataatcaat gccaacatgg cggtcatatt ggacatgagc
     7201 caatataaat gtacatatta tgatatagat acaacgtatg caatggccaa tagccaatat
     7261 tgatttatgc tatataacca atgaataata tggctaatgg ccaatattga gtcaacgcca
     7321 ctgtgcatga agctcgtaat tgttatccgc tcacaattaa gggcgacaca aaatttattc
     7381 taaatgataa taaatactga taacatctta tagtttgtat tatattttgt attatcgttg
     7441 acatgtataa ttttgatatc aaaaactgat tttcccttta ttattttcga gatttatttt
     7501 cttaattctc tttaacaaac tagaaatatt gtatatacaa aaaatcataa ataatagatg
     7561 aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac gtatcttatt taaagtgcgt
     7621 tgcttttttc tcatttataa ggttaaataa ttctcatata tcaagcaaag tgacaggcgc
     7681 ccttaaatat tctgacaaat gctctttccc taaactcccc ccataaaaaa acccgccgaa
     7741 gcgggttttt acgttatttg cggattaacg attactc
//
