LOCUS       sequence-304095-        9013 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 9013)
  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 9013)
  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-PIGR-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..9013
                     /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             4618..4653
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             4663..4686
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6346..6390
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6391..6417
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(6557..6574)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6563..6674
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6792..6812)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6797..7126
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6977..7112
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7039..7058
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7174..7221
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7240..7839
                     /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(7240..7259)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7736..7756
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7969..8090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8006..8025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8060..8079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8143..8934,8935..9003))
                     /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     8766..8785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9004..9013,1..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 cggccgccaa gtctcctatc
     2761 tttggccctg aggaagtgaa cagcgtcgag ggcaatagcg tgtccatcac ctgttactac
     2821 cctccaacca gcgtgaaccg gcacaccaga aagtactggt gtagacaggg cgccagaggc
     2881 ggctgtatca cactgattag cagcgagggc tacgtgtcct ctaagtacgc cggcagagcc
     2941 aacctgacca actttcccga gaatggcacc ttcgtggtca atatcgccca gctgagccag
     3001 gacgacagcg gcagatacaa gtgtggcctg ggcatcaata gcagaggcct gagcttcgac
     3061 gtgtccctgg aagtgtctca aggccctggc ctgctgaacg acaccaaggt gtacacagtg
     3121 gacctgggca gaaccgtgac catcaactgc cccttcaaga ccgagaacgc ccagaagaga
     3181 aagagcctgt acaagcagat cggactgtac cccgtgctgg tcatcgatag cagcggctac
     3241 gtgaacccca actacaccgg cagaatccgg ctggatattc aaggcaccgg ccagctgctg
     3301 ttcagcgtgg tcatcaatca gctgagactg agcgacgccg gccagtatct ttgtcaggcc
     3361 ggcgacgaca gcaacagcaa caagaagaat gccgacctcc aggtgctgaa gcctgagcct
     3421 gagctggtgt atgaggatct gagaggcagc gtgaccttcc actgtgctct gggacctgaa
     3481 gtggccaacg tggccaagtt tctgtgcaga cagagcagcg gcgagaactg cgacgtggtg
     3541 gttaataccc tgggaaagag agcccctgcc ttcgagggaa gaatcctgct gaatccccag
     3601 gacaaggacg gcagctttag cgtcgtgatc accggcctga gaaaagagga cgccggaaga
     3661 tatctgtgtg gcgcccattc cgatggacag ctgcaagagg gatctccaat tcaggcctgg
     3721 cagctgttcg tgaacgagga aagcacaatc cctagaagcc ccaccgtggt taagggcgtt
     3781 gcaggcggat ctgtggccgt gctgtgcccc tacaacagaa aagagagcaa gagcatcaag
     3841 tattggtgcc tgtgggaggg cgcccagaat ggaagatgtc ctctgctggt ggatagcgaa
     3901 ggctgggtca aagcccagta cgagggcaga ctgagcctgc tggaagaacc tggcaatggc
     3961 acattcaccg tgatcctgaa ccagctgaca agcagagatg ccggcttcta ctggtgcctg
     4021 accaatggcg ataccctgtg gcggaccacc gtggaaatca agatcatcga gggcgagccc
     4081 aatctgaagg tgcccggaaa tgtgacagcc gtgctgggag agacactgaa agtgccctgc
     4141 cactttcctt gcaagttcag cagctacgag aaatattggt gcaagtggaa caacaccggc
     4201 tgtcaggctc tgcctagcca ggatgaaggc cctagcaagg ccttcgtgaa ttgcgacgag
     4261 aactcccggc tggtgtctct gaccctgaat ctggtcacaa gagccgatga aggctggtat
     4321 tggtgtggcg tgaagcaggg ccacttttat ggcgagacag ccgccgtgta cgtggccgtg
     4381 gaagaaagaa aagccgccgg aagcagggat gtgtctctgg ccaaagctga tgccgctcct
     4441 gacgagaagg tgctggatag cggcttcaga gagatcgaga acaaggccat tcaggacccc
     4501 agactgttcg ccgaagaaaa ggccgtggcc gataccagag atcaggctga tggaagcaga
     4561 gcctccgtgg attctggcag ctctgaggaa caaggcggct ccagtagagg cgcgccggaa
     4621 gatcaagttg atccaagatt gattgatggc aagggcggat ctctggaggt gctgttccag
     4681 ggaccaggcg gaagcggcaa tcccggcggc gatctggccc cccagatgct gcgcgagctg
     4741 caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag
     4801 atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggtatgca gccctctaga
     4861 ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa ccgcgaggca
     4921 gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc caagaacctc
     4981 atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag gatcctaaaa
     5041 gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg cttcccatat
     5101 gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg agccacagcc
     5161 acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag tgcagccgcc
     5221 cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat gaatcgggcc
     5281 aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca cgcctcgcca
     5341 gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga cgtgcctgcc
     5401 tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa catggacatt
     5461 gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc agaccctgag
     5521 tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct ggtgcaggaa
     5581 tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct catgcaggct
     5641 tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga catgaaatac
     5701 gagatccacc gagactccac actggacccc tccctgatgg agatgacaga ggctgccctg
     5761 cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac
     5821 catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat gttcgacgac
     5881 gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct cgtcactgcc
     5941 gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc catcttcggg
     6001 ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata cggaaacggt
     6061 ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga gagcgggagc
     6121 cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc aggcgaggac
     6181 gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca ggagcagacc
     6241 ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg
     6301 gcgccccccg ccggcaccac cgacgccgcg cacccgggtg gcggaggact gaacgacatc
     6361 ttcgaggctc agaaaatcga atggcacgaa catcaccatc accatcacca tcaccattaa
     6421 ggccggcctg tagtcactgg cacaacgcgt tacccggtaa gccaatcggg tatacacggt
     6481 cgtcatactg cagacagggt tcttctactt tgcaagatag tcttgagtag taaaataaat
     6541 agatagagaa aatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6601 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6661 gaggaaattg catcacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6721 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6781 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     6841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     6901 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     6961 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     7021 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     7081 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     7141 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     7201 atcggcatag tataatacga caaggtgagg aactaaacca tgaccgagta caagcccacg
     7261 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     7321 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     7381 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     7441 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     7501 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     7561 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     7621 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     7681 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacatccgc gccccgcaac
     7741 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     7801 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac acgtgctacg agatttcgat
     7861 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     7921 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     7981 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     8041 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     8101 ataccgtcga cctctagcta gagcttggcg taatcatggt cattaccaat gcttaatcag
     8161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8221 cgtgtagata actacgatac gggagggctt accatctggc cccagcgctg cgatgatacc
     8281 gcgagaacca cgctcaccgg ctccggattt atcagcaata aaccagccag ccggaagggc
     8341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccatcgctac
     8461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     9001 catattcttc ctt
//
