LOCUS       sequence-304059-        8779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            COMP5AP-AviTag-9xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8779)
  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 8779)
  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-WFIKKN2-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     149..168
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      254..842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     996..1013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1111..1128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1112..1128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1584..1600
                     /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        1767..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2347..2371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2581..2599
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2581..2598
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2612..2631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4384..4419
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             4429..4452
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6112..6156
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6157..6183
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(6323..6340)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6329..6440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6558..6578)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6563..6892
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6743..6878
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6805..6824
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6940..6987
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7006..7605
                     /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(7006..7025)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7502..7522
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7735..7856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7772..7791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7826..7845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7909..8700,8701..8769))
                     /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     8532..8551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8770..8779,1..82))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
      121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
      181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
      241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
     1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
     1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
     1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
     1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
     1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
     1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
     1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
     1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
     1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
     1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
     1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
     2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
     2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
     2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgcctt gcctcctatc
     2761 agatattctc acgccggcat ctgccccaac gacatgaacc ctaatctgtg ggtcgacgcc
     2821 cagagcacct gtcggagaga gtgcgagaca gaccaagagt gcgaaaccta cgagaagtgc
     2881 tgccccaatg tgtgcggcac caagtcttgt gtggccgcca gatatatgga cgtgaagggc
     2941 aagaaaggcc ccgtgggcat gcctaaagag gccacttgcg accacttcat gtgcctgcag
     3001 cagggcagcg agtgcgacat ttgggatgga cagcctgtgt gcaagtgcaa ggacagatgc
     3061 gagaaagagc ccagctttac ctgcgcctct gacggcctga cctactacaa ccggtgctac
     3121 atggatgccg aggcctgctc taagggcatc acactggccg tggtcacctg tagataccac
     3181 ttcacctggc ctaacacaag ccctccacca cctgagacaa caatgcaccc caccaccgcc
     3241 tctccagaga cacctgaact ggatatggcc gctcctgctc tgctgaacaa ccctgtgcac
     3301 cagagcgtga ccatgggcga gacagtgtcc ttcctgtgtg acgtcgtggg cagacccaga
     3361 cctgagatca catgggagaa gcagctggaa gatcgcgaga acgtggtcat gaggcccaat
     3421 cacgtgcggg gcaatgtggt ggtcaccaat atcgcccagc tggtcatcta caatgctcag
     3481 ctgcaggacg ccggaatcta cacctgtacc gccagaaatg tggccggcgt gctgagagcc
     3541 gattttccac tgtctgttgt gcggggacat caggccgctg ccacatctga gtctagcccc
     3601 aatggcacag cctttccagc cgccgagtgt ctgaagcctc ctgactctga agattgcggc
     3661 gaggaacaga cccggtggca ctttgatgct caggccaaca actgcctgac cttcaccttc
     3721 ggccactgcc acagaaacct gaaccacttc gagacatatg aggcctgtat gctggcctgc
     3781 atgtctggac ctctggccgc ttgttctctg ccagctctgc agggcccatg caaggcctat
     3841 gctcctagat gggcctacaa cagccagacc ggacagtgcc agagctttgt gtacggcggc
     3901 tgtgaaggca acggcaacaa cttcgagtcc agagaggcct gcgaggaaag ctgcccattt
     3961 cctagaggca accagcggtg cagagcctgc aagcccagac agaagctggt cacaagcttt
     4021 tgtcggagcg acttcgtgat cctgggcaga gtgtccgagc tgaccgagga acctgattct
     4081 ggcagagccc tggtcaccgt ggatgaggtg ctgaaggatg agaagatggg actgaagttc
     4141 ctgggccaag agcctctgga agtgacactg ctgcatgtgg actgggcctg tccttgtcct
     4201 aacgtgaccg tgtccgagat gcccctgatc atcatgggcg aagtggatgg cggaatggcc
     4261 atgctgaggc ccgattcttt tgtgggcgcc agcagcgcca gaagagtgcg gaagctgagg
     4321 gaagtgatgc acaagaaaac ctgcgacgtg ctgaaagagt ttctgggcct gcacggcgcg
     4381 ccggaagatc aagttgatcc aagattgatt gatggcaagg gcggatctct ggaggtgctg
     4441 ttccagggac caggcggaag cggcaatccc ggcggcgatc tggcccccca gatgctgcgc
     4501 gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg
     4561 aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg tatgcagccc
     4621 tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt ctggaaccgc
     4681 gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac agccgccaag
     4741 aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc tgccaggatc
     4801 ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat ggaccgcttc
     4861 ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga cagtggagcc
     4921 acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg cttgagtgca
     4981 gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc cgtgatgaat
     5041 cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt gcagcacgcc
     5101 tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga cgccgacgtg
     5161 cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat ctccaacatg
     5221 gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg aaccccagac
     5281 cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa gaatctggtg
     5341 caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg
     5401 caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc tggagacatg
     5461 aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat gacagaggct
     5521 gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc
     5581 atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac gatcatgttc
     5641 gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct gagcctcgtc
     5701 actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg gagctccatc
     5761 ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct cctatacgga
     5821 aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga gagcgagagc
     5881 gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac ccacgcaggc
     5941 gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag
     6001 cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta caccgcctgc
     6061 gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggtggcgg aggactgaac
     6121 gacatcttcg aggctcagaa aatcgaatgg cacgaacatc accatcacca tcaccatcac
     6181 cattaaggcc ggcctgtagt cactggcaca acgcgttacc cggtaagcca atcgggtata
     6241 cacggtcgtc atactgcaga cagggttctt ctactttgca agatagtctt gagtagtaaa
     6301 ataaatagat agagaaaatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6421 taaaatgagg aaattgcatc acaacactca accctatctc ggtctattct tttgatttat
     6481 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6541 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6601 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     6661 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     6721 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     6781 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     6841 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     6901 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     6961 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatgac cgagtacaag
     7021 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     7081 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     7141 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     7201 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     7261 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     7321 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     7381 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     7441 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac atccgcgccc
     7501 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     7561 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacacgt gctacgagat
     7621 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     7681 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     7741 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7801 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7861 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcatt accaatgctt
     7921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gcgctgcgat
     8041 gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc agccagccgg
     8101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8221 cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8761 aatactcata ttcttcctt
//
