LOCUS       sequence-420991-        8245 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 8245)
  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 8245)
  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-AMIGO3-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(85..176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     218..237
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     243..262
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      348..936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     837..856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1090..1107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1205..1222
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1206..1222
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1678..1694
                     /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        1861..2240
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2241..2444
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2395..2415
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2441..2465
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2675..2693
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2675..2692
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2706..2725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3944..3979
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             3989..4012
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5672..5716
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5717..5743
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(5883..5900)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5889..6000
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6118..6138)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6123..6452
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6303..6438
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6500..6547
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6566..7165
                     /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(6566..6585)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7062..7082
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7295..7416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7332..7351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7386..7405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7469..8245,1..15,16..84))
                     /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     8092..8111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
       61 gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa gcatttatca
      121 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      181 ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat ttatacctga
      241 atatggctca taacacccct tgctcatgac caaaatccct taacgtgagt tacgcgcgcg
      301 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      361 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      421 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      481 ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa gaactctgta
      541 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      601 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      661 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      721 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      781 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      841 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      901 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      961 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1021 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1081 accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga actgccaggc
     1141 atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca aactctttct
     1201 gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct gataacgagt
     1261 aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt
     1321 ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct tgatatatga
     1381 gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt
     1441 tttcgattga tgaacaccta taattaaact attcatctat tatttatgat tttttgtata
     1501 tacaatattt ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa taataaaggg
     1561 aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat ataatacaaa
     1621 ctataagatg ttatcagtat ttattatcat ttagaataaa ttttgtgtcg cccttaattg
     1681 tgagcggata acaattacga gcttcatgca cagtggcgtt gactcaatat tggccattag
     1741 ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     1801 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
     1861 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     1921 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     1981 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2041 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     2101 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     2161 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     2221 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     2281 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     2341 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     2401 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
     2461 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     2521 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
     2581 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
     2641 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
     2701 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
     2761 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
     2821 cctggtgttc gctgcggccg ccacacctga ttctgagggc tttccacctc gggctctgca
     2881 caactgcccc tacaagtgta tctgtgccgc cgacctgctg agctgtacag gactgggact
     2941 gcaggatgtg cctgccgaac ttcctgccgc tacagccgat ctggacctgt ctcataacgc
     3001 cctgcagaga ctgaggcctg gatggctggc tcctctgttt cagctgagag ccctgcacct
     3061 ggaccacaac gaactggatg ctctcggcag aggcgtgttc gtgaatgcct ctggactgag
     3121 actgctggac ctgagcagca acaccctgag agcactgggc agacacgatc tggatggact
     3181 gggagccctg gaaaagctgc tgctgttcaa caaccggctg gtgcatctgg acgagcacgc
     3241 ctttcatgga ctgagggctc tgagccacct gtacctgggc tgtaatgagc tggcctcctt
     3301 cagcttcgac catctgcacg gactgagcgc cacacatctg ctgacactgg atctgtccag
     3361 caacagactg ggccacatca gcgtgccaga actggcagct ctgcctgcct tcctgaagaa
     3421 cggcctgtat ctgcataaca accctctgcc ttgcgactgc cggctgtacc atctgctgca
     3481 aagatggcat cagaggggcc tgtctgccgt gcgagatttc gccagagaat acgtgtgcct
     3541 ggccttcaag gtgcccgcca gtagagtgcg gttcttccag cacagccggg tgttcgagaa
     3601 ttgctctagc gcccctgctc tgggcctcga aagaccagag gaacacctgt atgccctcgt
     3661 gggcagaagc ctgagactgt actgcaatac ctccgtgcct gccatgagaa tcgcctgggt
     3721 ttccccacag caagagctgc ttagagcccc tggaagcaga gatggctcta ttgccgtgct
     3781 ggccgatggc tctctggcca tcggaaatgt gcaagagcag cacgccggcc tgtttgtgtg
     3841 tctggcaaca ggacctcggc tgcaccacaa tcagacccac gagtacaacg tgtccgtgca
     3901 cttccccaga cctgagcctg aggccttcaa cacaggcgcg ccggaagatc aagttgatcc
     3961 aagattgatt gatggcaagg gcggatctct ggaggtgctg ttccagggac caggcggaag
     4021 cggcaatccc ggcggcgatc tggcccccca gatgctgcgc gagctgcagg agaccaatgc
     4081 cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg aaggagatca ccttcctgaa
     4141 gaacaccgtg atggagtgcg acgcctgcgg tatgcagccc tctagaggct ccggctccat
     4201 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     4261 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     4321 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     4381 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     4441 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     4501 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     4561 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     4621 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     4681 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     4741 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     4801 tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     4861 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     4921 ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     4981 tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     5041 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     5101 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     5161 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     5221 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     5281 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     5341 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     5401 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     5461 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     5521 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     5581 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     5641 caccaccgac gccgcgcacc cgggtggcgg aggactgaac gacatcttcg aggctcagaa
     5701 aatcgaatgg cacgaacatc accatcacca tcaccatcac cattaaggcc ggcctgtagt
     5761 cactggcaca acgcgttacc cggtaagcca atcgggtata cacggtcgtc atactgcaga
     5821 cagggttctt ctactttgca agatagtctt gagtagtaaa ataaatagat agagaaaatc
     5881 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     5941 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6001 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6061 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     6121 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6181 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6241 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6301 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6361 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     6421 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     6481 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     6541 atacgacaag gtgaggaact aaaccatgac cgagtacaag cccacggtgc gcctcgccac
     6601 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     6661 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     6721 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     6781 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     6841 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     6901 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     6961 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     7021 gcgcgccggg gtgcccgcct tcctggagac atccgcgccc cgcaacctcc ccttctacga
     7081 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     7141 catgacccgc aagcccggtg cctgacacgt gctacgagat ttcgattcca ccgccgcctt
     7201 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     7261 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     7321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     7381 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     7441 tagctagagc ttggcgtaat catggtcatt accaatgctt aatcagtgag gcacctatct
     7501 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7561 cgatacggga gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct
     7621 caccggctcc ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7681 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7741 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt
     7801 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7861 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7921 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7981 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8041 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8101 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8161 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8221 gatcttcagc atcttttact ttcac
//
