LOCUS       sequence-385762-        8182 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 8182)
  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 8182)
  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-CADM2-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8182
                     /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             2021..2056
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2066..2089
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3749..3793
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3794..3820
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3960..3977)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3966..4077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4195..4215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4200..4529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4380..4515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4442..4461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4577..4624
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4643..5242
                     /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(4643..4662)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5139..5159
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5372..5493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5409..5428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5463..5482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5546..6337,6338..6406))
                     /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     6169..6188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6407..6498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6540..6559
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6565..6584
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6670..7258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7159..7178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7412..7429
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7527..7544
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7528..7544
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8000..8016
                     /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 cccaatttcc tctgacacag aacgtgacag tggtggaagg
     1021 cggcaccgcc atcctgacat gcagagtgga ccagaacgac aacaccagcc tgcagtggtc
     1081 caatcctgct cagcagaccc tgtacttcga cgacaagaag gccctgcggg acaacagaat
     1141 cgaacttgtg cgggcctctt ggcacgagct gagcatctct gtgtccgatg tgtccctgag
     1201 cgacgagggc cagtacacct gtagcctgtt caccatgcct gtgaaaacca gcaaggccta
     1261 cctgacagtg ctgggcgtgc cagagaagcc tcagatctct ggctttagca gccccgtgat
     1321 ggaaggcgat ctgatgcagc tgacctgcaa gacatctggc tctaagcctg ccgccgacat
     1381 ccggtggttc aagaatgaca aagagatcaa ggacgtcaag tacctgaaag aagaggacgc
     1441 caacagaaag accttcaccg tgtctagcac cctggacttc agagtggata gaagcgacga
     1501 tggcgtggcc gtgatctgta gagtggatca cgagagcctg aacgccacac ctcaggtggc
     1561 aatgcaggtc ctggaaatcc actacacccc tagcgtgaag atcatcccca gcacaccctt
     1621 tccacaagag ggccagcctc tgatcctgac ctgtgaaagc aagggcaagc cactgcctga
     1681 gcctgtgctg tggacaaaag atggcggaga gctgcccgat cctgacagaa tggtggtgtc
     1741 cggcagagag ctgaatatcc tgttcctgaa caagaccgac aacggcacct acagatgcga
     1801 ggccacaaat accatcggcc agagcagcgc cgagtacgtg ctgattgtgc acgatgtgcc
     1861 caacacactg ctgcccacca caatcatccc tagcctgacc accgccaccg tgacaacaac
     1921 agtggccatc acaacaagcc ctaccaccag cgccaccacc agcagcatca gagatcctaa
     1981 tgctctggct ggccagaacg gccccgatca tggcgcgccg gaagatcaag ttgatccaag
     2041 attgattgat ggcaagggcg gatctctgga ggtgctgttc cagggaccag gcggaagcgg
     2101 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     2161 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     2221 caccgtgatg gagtgcgacg cctgcggtat gcagccctct agaggctccg gctccatcat
     2281 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     2341 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     2401 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     2461 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     2521 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     2581 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     2641 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     2701 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     2761 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     2821 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     2881 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     2941 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     3001 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     3061 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     3121 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     3181 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     3241 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     3301 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     3361 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     3421 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     3481 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     3541 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     3601 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     3661 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     3721 caccgacgcc gcgcacccgg gtggcggagg actgaacgac atcttcgagg ctcagaaaat
     3781 cgaatggcac gaacatcacc atcaccatca ccatcaccat taaggccggc ctgtagtcac
     3841 tggcacaacg cgttacccgg taagccaatc gggtatacac ggtcgtcata ctgcagacag
     3901 ggttcttcta ctttgcaaga tagtcttgag tagtaaaata aatagataga gaaaatcagc
     3961 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4021 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcaca
     4081 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4141 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4201 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4261 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4321 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4381 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4441 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4501 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4561 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     4621 cgacaaggtg aggaactaaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4681 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4741 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4801 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4861 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4921 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4981 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5041 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5101 cgccggggtg cccgccttcc tggagacatc cgcgccccgc aacctcccct tctacgagcg
     5161 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5221 gacccgcaag cccggtgcct gacacgtgct acgagatttc gattccaccg ccgccttcta
     5281 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5341 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5521 ctagagcttg gcgtaatcat ggtcattacc aatgcttaat cagtgaggca cctatctcag
     5581 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5641 tacgggaggg cttaccatct ggccccagcg ctgcgatgat accgcgagaa ccacgctcac
     5701 cggctccgga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5761 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5821 gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac
     5881 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5941 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6001 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6061 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6121 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6181 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6241 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6301 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6361 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatattc ttcctttttc
     6421 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6481 tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa ttaaccaatt ctgaacatta
     6541 tcgcgagccc atttatacct gaatatggct cataacaccc cttgctcatg accaaaatcc
     6601 cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6661 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6721 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6781 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagc
     6841 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6901 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6961 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7021 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7081 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7141 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7201 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7261 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7321 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7381 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg
     7441 cgagagtagg gaactgccag gcatcaaact aagcagaagg cccctgacgg atggcctttt
     7501 tgcgtttcta caaactcttt ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc
     7561 ctgggcggtt ctgataacga gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg
     7621 cgggtttttt tatgggggga gtttagggaa agagcatttg tcagaatatt taagggcgcc
     7681 tgtcactttg cttgatatat gagaattatt taaccttata aatgagaaaa aagcaacgca
     7741 ctttaaataa gatacgttgc tttttcgatt gatgaacacc tataattaaa ctattcatct
     7801 attatttatg attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat
     7861 aaatctcgaa aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg
     7921 ataatacaaa atataataca aactataaga tgttatcagt atttattatc atttagaata
     7981 aattttgtgt cgcccttaat tgtgagcgga taacaattac gagcttcatg cacagtggcg
     8041 ttgactcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
     8101 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     8161 tgtccaatat gaccgccatg tt
//
