LOCUS       sequence-405785-        8143 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 8143)
  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 8143)
  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-CADM1-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1982..2017
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2027..2050
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3710..3754
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3755..3781
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3921..3938)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3927..4038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4156..4176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4161..4490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4341..4476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4403..4422
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4538..4585
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4604..5203
                     /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(4604..4623)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5100..5120
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5333..5454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5370..5389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5424..5443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5507..6298,6299..6367))
                     /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     6130..6149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6368..6459)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6501..6520
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6526..6545
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6631..7219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7120..7139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7373..7390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7488..7505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7489..7505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7961..7977
                     /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)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg cccagaatct gttcaccaag gacgtgacag tgatcgaggg
     1021 cgaagtggcc accatcagct gccaagtgaa caagagcgac gacagcgtga tccagctgct
     1081 gaaccccaac cggcagacca tctacttcag agacttcaga cccctgaagg acagccggtt
     1141 ccagctcctg aatttcagca gcagcgagct gaaggtgtcc ctgaccaatg tgtccatcag
     1201 cgacgagggc agatacttct gccagctgta caccgatcct ccacaagaga gctacaccac
     1261 catcaccgtg ctggtgcctc cacggaacct gatgatcgac atccagaaag acaccgccgt
     1321 ggaaggcgaa gagatcgaag tgaattgcac cgccatggcc agcaagcccg ccaccacaat
     1381 cagatggttc aagggcaaca ccgagctgaa aggcaagagc gaggtggaag agtggtccga
     1441 catgtacacc gtgaccagcc agctgatgct gaaagtgcac aaagaggatg acggcgtccc
     1501 cgtgatctgt caggtggaac atcctgccgt gaccggcaat ctgcagaccc agagatacct
     1561 ggaagtgcag tacaagcccc aggtgcacat ccagatgaca taccctctgc aaggcctgac
     1621 aagagagggc gacgctctgg aactgacatg cgaggccatt ggaaagcccc agcctgtgat
     1681 ggtcacctgg gtccgagttg acgatgagat gcctcagcat gccgtgctga gcggccctaa
     1741 cctgttcatc aacaacctga acaagacgga caacggcacc taccgctgcg aggcctctaa
     1801 tatcgtgggc aaagcccaca gcgactacat gctgtacgtg tacgaccctc ctaccaccat
     1861 tcctccacca accaccacca caaccactac taccacaacg acaaccacca tcctgaccat
     1921 catcaccgac agcagagccg gcgaggaagg atctatcaga gccgtggatc atggcgcgcc
     1981 ggaagatcaa gttgatccaa gattgattga tggcaagggc ggatctctgg aggtgctgtt
     2041 ccagggacca ggcggaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     2101 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     2161 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggta tgcagccctc
     2221 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     2281 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     2341 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     2401 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     2461 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     2521 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     2581 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     2641 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     2701 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     2761 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     2821 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     2881 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2941 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     3001 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     3061 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     3121 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     3181 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     3241 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     3301 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     3361 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     3421 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     3481 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     3541 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     3601 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     3661 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggtggcggag gactgaacga
     3721 catcttcgag gctcagaaaa tcgaatggca cgaacatcac catcaccatc accatcacca
     3781 ttaaggccgg cctgtagtca ctggcacaac gcgttacccg gtaagccaat cgggtataca
     3841 cggtcgtcat actgcagaca gggttcttct actttgcaag atagtcttga gtagtaaaat
     3901 aaatagatag agaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3961 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4021 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     4081 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4141 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4321 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4381 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4441 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4501 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     4561 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc
     4621 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4681 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4741 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4801 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4861 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4921 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4981 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5041 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg
     5101 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5161 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt
     5221 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5281 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5341 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5401 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5461 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa
     5521 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5581 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga
     5641 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa
     5701 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5761 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     5821 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5881 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5941 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6001 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6061 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6121 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6181 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6241 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6301 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6361 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6421 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca
     6481 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc
     6541 ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     6601 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6661 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6721 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6781 tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct
     6841 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6901 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6961 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7021 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7081 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7141 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7201 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7261 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7321 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7381 agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag
     7441 gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg
     7501 ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa
     7561 taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt
     7621 gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat
     7681 aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac
     7741 ctataattaa actattcatc tattatttat gattttttgt atatacaata tttctagttt
     7801 gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata
     7861 tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag atgttatcag
     7921 tatttattat catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta
     7981 cgagcttcat gcacagtggc gttgactcaa tattggccat tagccatatt attcattggt
     8041 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     8101 atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
