LOCUS       sequence-420998-        8188 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 8188)
  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 8188)
  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-AMIGO1-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1094..1138
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1139..1165
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(1305..1322)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1311..1422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1540..1560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1545..1874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1725..1860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1787..1806
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1922..1969
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1988..2587
                     /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(1988..2007)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2484..2504
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2717..2838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2754..2773)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2808..2827
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(2891..3682,3683..3751))
                     /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     3514..3533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3752..3843)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3885..3904
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3910..3929
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4015..4603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4504..4523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4757..4774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4872..4889
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4873..4889
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5345..5361
                     /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        5528..5907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5908..6111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6062..6082
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6108..6132
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        6342..6360
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6342..6359
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6373..6392
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             7554..7589
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             7599..7622
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
       61 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
      121 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
      181 tacgcagctc atctccaaca tggacattga cgtgatccta ggtggaggcc gaaagtacat
      241 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
      301 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
      361 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
      421 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
      481 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
      541 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
      601 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
      661 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
      721 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
      781 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
      841 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
      901 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
      961 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     1021 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     1081 cccgggtggc ggaggactga acgacatctt cgaggctcag aaaatcgaat ggcacgaaca
     1141 tcaccatcac catcaccatc accattaagg ccggcctgta gtcactggca caacgcgtta
     1201 cccggtaagc caatcgggta tacacggtcg tcatactgca gacagggttc ttctactttg
     1261 caagatagtc ttgagtagta aaataaatag atagagaaaa tcagcctcga ctgtgccttc
     1321 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1381 cactcccact gtcctttcct aataaaatga ggaaattgca tcacaacact caaccctatc
     1441 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1501 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     1561 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1621 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1681 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     1741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     1801 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     1861 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     1921 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     1981 ctaaaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     2041 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     2101 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     2161 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     2221 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     2281 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     2341 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     2401 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     2461 cttcctggag acatccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     2521 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     2581 tgcctgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     2641 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     2701 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2761 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2821 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     2881 atcatggtca ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2941 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3001 catctggccc cagcgctgcg atgataccgc gagaaccacg ctcaccggct ccggatttat
     3061 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3121 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3181 gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3241 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3301 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3361 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3421 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3481 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3541 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3601 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3661 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3721 taagggcgac acggaaatgt tgaatactca tattcttcct ttttcaatat tattgaagca
     3781 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3841 aaataggggt cagtgttaca accaattaac caattctgaa cattatcgcg agcccattta
     3901 tacctgaata tggctcataa caccccttgc tcatgaccaa aatcccttaa cgtgagttac
     3961 gcgcgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4021 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4081 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4141 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttagcccacc acttcaagaa
     4201 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4261 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4321 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4381 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4441 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4501 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4561 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4621 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4681 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4741 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaaggcgaga gtagggaact
     4801 gccaggcatc aaactaagca gaaggcccct gacggatggc ctttttgcgt ttctacaaac
     4861 tctttctgtg ttgtaaaacg acggccagtc ttaagctcgg gccccctggg cggttctgat
     4921 aacgagtaat cgttaatccg caaataacgt aaaaacccgc ttcggcgggt ttttttatgg
     4981 ggggagttta gggaaagagc atttgtcaga atatttaagg gcgcctgtca ctttgcttga
     5041 tatatgagaa ttatttaacc ttataaatga gaaaaaagca acgcacttta aataagatac
     5101 gttgcttttt cgattgatga acacctataa ttaaactatt catctattat ttatgatttt
     5161 ttgtatatac aatatttcta gtttgttaaa gagaattaag aaaataaatc tcgaaaataa
     5221 taaagggaaa atcagttttt gatatcaaaa ttatacatgt caacgataat acaaaatata
     5281 atacaaacta taagatgtta tcagtattta ttatcattta gaataaattt tgtgtcgccc
     5341 ttaattgtga gcggataaca attacgagct tcatgcacag tggcgttgac tcaatattgg
     5401 ccattagcca tattattcat tggttatata gcataaatca atattggcta ttggccattg
     5461 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     5521 ccatgttggc attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     5581 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     5641 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     5701 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     5761 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     5821 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     5881 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     5941 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     6001 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     6061 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     6121 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     6181 gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa cgcggattcc ccgtgccaag
     6241 agtgacgtaa gtaccgccta tagagtctat aggcccaccc ccttggcttc gttagaacgc
     6301 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactataga
     6361 ataacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa ctgcacctcg
     6421 gttctatcga aaacgcgcct gctagcgcca ccatgaagac catcattgcc ctgagctaca
     6481 tcttctgcct ggtgttcgct gcggccgccg gtagagctgt tgtttcttgt cctgctgcct
     6541 gcctgtgcgc cagcaatatc ctgagctgta gcaagcagca gctgcccaat gtgcctcaca
     6601 gcctgccatc ttacacagcc ctgctggacc tgagccacaa caacctgagc agactgagag
     6661 ccgagtggac ccctacaaga ctgacacagc tgcactccct gctgctgtcc cacaatcacc
     6721 tgaacttcat cagcagcgag gccttctctc ccgtgcctaa cctgagatac ctggacctgt
     6781 ccagcaacca gctgagaacc ctggacgagt tcctgttcag cgacctccag gtgctggaag
     6841 tgctgctgct ctacaacaac cacatcatgg ccgtggacag atgcgccttc gatgatatgg
     6901 cccagctgca gaagctgtat ctgagccaga accagatcag cagattccca ctggaactgg
     6961 tcaaagaggg cgccaagctg cctaagctga cactgctgga tctgtcctcc aacaagctga
     7021 agaatctgcc cctgcctgac ctgcagaaac tgcccgcctg gatcaagaat ggcctgtacc
     7081 tgcacaacaa tcctctgaac tgcgactgcg agctgtacca gctgttcagc cactggcagt
     7141 acagacagct gagcagcgtg atggacttcc aagaggacct gtactgcatg aacagcaaga
     7201 aactgcacaa cgtgttcaac ctgtccttcc tgaattgcgg cgagtacaaa gagagagcct
     7261 gggaagccca cctgggcgac acactgatca tcaagtgcga caccaaacag cagggcatga
     7321 ccaaagtgtg ggtcacccct agcaacgagc gcgtgctgga tgaagtgacc aatggcaccg
     7381 tgtccgtgtc caaggatggc agccttctgt tccagcaagt gcaggttgag gacggcggcg
     7441 tgtacacatg ttatgccatg ggcgagacat tcaacgagac actgagcgtg gaactgaagg
     7501 tgcacaactt cacactgcac ggccaccacg acaccctgaa cacaggcgcg ccggaagatc
     7561 aagttgatcc aagattgatt gatggcaagg gcggatctct ggaggtgctg ttccagggac
     7621 caggcggaag cggcaatccc ggcggcgatc tggcccccca gatgctgcgc gagctgcagg
     7681 agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg aaggagatca
     7741 ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg tatgcagccc tctagaggct
     7801 ccggctccat catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg
     7861 aggccctggg tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca
     7921 tcttcctggg cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc
     7981 agaagaagga caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg
     8041 ctctgtccaa gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg
     8101 cctacctgtg cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct
     8161 ttaaccagtg caacacgaca cgcggcaa
//
