LOCUS       sequence-383697-        8935 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 8935)
  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 8935)
  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-FAS-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8935
                     /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     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        3105..3484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3485..3688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3639..3659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3685..3709
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3919..3937
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3919..3936
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     3950..3969
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4540..4575
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             4585..4608
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6268..6312
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6313..6339
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(6479..6496)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6485..6596
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6714..6734)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6719..7048
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6899..7034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6961..6980
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7096..7143
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7162..7761
                     /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(7162..7181)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7658..7678
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7891..8012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7928..7947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7982..8001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8065..8856,8857..8925))
                     /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     8688..8707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8926..8935,1..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 gctgatgaat cccctaatga ttttggtaaa aatcattaag ttaaggtgga tacacatctt
     1681 gtcatatgat caaatggttt cgcgaaaaat caataatcag acaacaagat gtgcgaactc
     1741 gatattttac acgactctct ttaccaattc tgccccgaat tacacttaaa acgactcaac
     1801 agcttaacgt tggcttgcca cgcattactt gactgtaaaa ctctcactct taccgaactt
     1861 ggccgtaacc tgccaaccaa agcgagaaca aaacataaca tcaaacgaat cgaccgattg
     1921 ttaggtaatc gtcacctcca caaagagcga ctcgctgtat accgttggca tgctagcttt
     1981 atctgttcgg gcaatacgat gcccattgta cttgttgact ggtctgatat tcgtgagcaa
     2041 aaacgactta tggtattgcg agcttcagtc gcactacacg gtcgttctgt tactctttat
     2101 gagaaagcgt tcccgctttc agagcaatgt tcaaagaaag ctcatgacca atttctagcc
     2161 gaccttgcga gcattctacc gagtaacacc acaccgctca ttgtcagtga tgctggcttt
     2221 aaagtgccat ggtataaatc cgttgagaag ctgggttggt actggttaag tcgagtaaga
     2281 ggaaaagtac aatatgcaga cctaggagcg gaaaactgga aacctatcag caacttacat
     2341 gatatgtcat ctagtcactc aaagacttta ggctataaga ggctgactaa aagcaatcca
     2401 atctcatgcc aaattctatt gtataaatct cgctctaaag gccgaaaaaa tcagcgctcg
     2461 acacggactc attgtcacca cccgtcacct aaaatctact cagcgtcggc aaaggagcca
     2521 tgggttctag caactaactt acctgttgaa attcgaacac ccaaacaact tgttaatatc
     2581 tattcgaagc gaatgcagat tgaagaaacc ttccgagact tgaaaagtcc tgcctacgga
     2641 ctaggcctac gccatagccg aacgagcagc tcagagcgtt ttgatatcat gctgctaatc
     2701 gccctgatgc ttcaactaac atgttggctt gcgggcgttc atgctcagaa acaaggttgg
     2761 gacaagcact tccaggctaa cacagtcaga aatcgaaacg tactctcaac agttcgctta
     2821 ggcatggaag ttttgcggca ttctggctac acaataacaa gggaagactt actcgtggct
     2881 gcaaccctac tagctcaaaa tttattcaca catggttacg ctttggggaa attatgaggg
     2941 gatctctcag tgcacagtgg cgttgactca atattggcca ttagccatat tattcattgg
     3001 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
     3061 tatgtacatt tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac
     3121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     3181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     3241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     3301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     3541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     3601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     3661 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
     3721 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
     3781 ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag
     3841 agtctatagg cccaccccct tggcttcgtt agaacgcggc tacaattaat acataacctt
     3901 atgtatcata cacatacgat ttaggtgaca ctatagaata acatccactt tgcctttctc
     3961 tccacaggtg tccactccca ggtccaactg cacctcggtt ctatcgaaaa cgcgcctgct
     4021 agcgccacca tgaagaccat cattgccctg agctacatct tctgcctggt gttcgctgcg
     4081 gccgcccaag tgaccgacat caatagcaaa ggcctggaac tgagaaagac cgtgaccacc
     4141 gtggaaaccc agaacctgga aggactgcac cacgacggcc agttctgtca caaaccttgt
     4201 ccacctggcg agcggaaggc cagagattgc acagtgaatg gcgacgagcc cgattgcgtg
     4261 ccctgtcaag agggcaaaga gtacaccgac aaggcccact tcagcagcaa gtgcagacgg
     4321 tgcagactgt gtgatgaagg ccacggcctg gaagtggaaa tcaactgcac ccggacacag
     4381 aacaccaagt gccggtgcaa gcccaacttc ttctgcaaca gcaccgtgtg cgagcactgc
     4441 gacccttgca caaagtgcga acacggcatc atcaaagagt gcaccctgac ctccaatacc
     4501 aagtgcaaag aggaaggcag ccggtccaac ggcgcgccgg aagatcaagt tgatccaaga
     4561 ttgattgatg gcaagggcgg atctctggag gtgctgttcc agggaccagg cggaagcggc
     4621 aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac caatgccgcc
     4681 ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt cctgaagaac
     4741 accgtgatgg agtgcgacgc ctgcggtatg cagccctcta gaggctccgg ctccatcatc
     4801 ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc
     4861 gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt cctgggcgat
     4921 gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa gaaggacaaa
     4981 ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct gtccaagaca
     5041 tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta cctgtgcggg
     5101 gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa ccagtgcaac
     5161 acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc agggaagtca
     5221 gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac ctacgcccac
     5281 acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg
     5341 tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat cctaggtgga
     5401 ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga tgactacagc
     5461 caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc gaagcgccag
     5521 ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga cccgtctgtg
     5581 acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca ccgagactcc
     5641 acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct gagcaggaac
     5701 ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca tcatgaaagc
     5761 agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga gagggcgggc
     5821 cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc
     5881 tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc tggcaaggcc
     5941 cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta tgtgctcaag
     6001 gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta tcggcagcag
     6061 tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt gttcgcgcgc
     6121 ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc gcacgtcatg
     6181 gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc cgccggcacc
     6241 accgacgccg cgcacccggg tggcggagga ctgaacgaca tcttcgaggc tcagaaaatc
     6301 gaatggcacg aacatcacca tcaccatcac catcaccatt aaggccggcc tgtagtcact
     6361 ggcacaacgc gttacccggt aagccaatcg ggtatacacg gtcgtcatac tgcagacagg
     6421 gttcttctac tttgcaagat agtcttgagt agtaaaataa atagatagag aaaatcagcc
     6481 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     6541 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcacaa
     6601 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     6661 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     6721 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6781 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     6841 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6901 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6961 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     7021 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     7081 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     7141 gacaaggtga ggaactaaac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     7201 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     7261 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     7321 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     7381 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     7441 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     7501 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     7561 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     7621 gccggggtgc ccgccttcct ggagacatcc gcgccccgca acctcccctt ctacgagcgg
     7681 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     7741 acccgcaagc ccggtgcctg acacgtgcta cgagatttcg attccaccgc cgccttctat
     7801 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     7861 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     7921 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7981 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     8041 tagagcttgg cgtaatcatg gtcattacca atgcttaatc agtgaggcac ctatctcagc
     8101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8161 acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac cacgctcacc
     8221 ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8341 ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg tggtgtcacg
     8401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct tcctt
//
