LOCUS       sequence-417399-        8986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8986)
  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 8986)
  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-KIRREL3-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8986
                     /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             3224..3268
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3269..3286
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3426..3443)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3432..3543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3661..3681)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3666..3995
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3846..3981
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3908..3927
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4043..4090
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4109..4708
                     /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(4109..4128)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4605..4625
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4838..4959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4875..4894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4929..4948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5012..5803,5804..5872))
                     /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     5635..5654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5873..5964)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6006..6025
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6031..6050
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6136..6724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6625..6644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6878..6895
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8331..8348
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8332..8348
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8804..8820
                     /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 ccttgcaaaa aaggggctgt tgtctggtgc tgggctacat
     1021 ggccaaggac aagttccggc ggatgaacga aggccaggtg tacagcttta gccagcagcc
     1081 tcaggaccag gtggtggtgt ctggacagcc tgtgacactg ctgtgcgcta tccctgagta
     1141 cgacggcttc gtgctgtgga tcaaggatgg actggccctc ggcgtgggca gagatctgag
     1201 tagctaccct cagtacctgg tcgtgggcaa tcacctgtct ggcgagcacc acctgaagat
     1261 cctgagagcc gagctgcagg acgatgccgt gtatgagtgt caggccatcc aggccgccat
     1321 cagaagcaga cctgctagac tgacagtgct ggtgcctcct gacgaccctg tgattcttgg
     1381 cggccctgtg atctctctga gagctggcga tcccctgaac ctgacctgcc acgccgataa
     1441 tgctaagcct gccgccagca tcatctggct gagaaagggc gaagtgatca acggcgccac
     1501 ctacagcaag accctgctga gagatggcaa gagagagagc atcgtcagca ccctgttcat
     1561 cagccctggc gacgtggaaa acggccagtc tatcgtgtgc cgggccacca acaaagccat
     1621 tccaggcggc aaagaaacca gcgtgaccat cgacattcag cacccacctc tggtcaacct
     1681 gagcgtggaa cctcagcctg tgctggaaga taacgtggtc accttccact gcagcgccaa
     1741 ggccaatcct gccgtgacac agtacagatg ggccaagaga ggccagatca tcaaagaggc
     1801 cagcggcgag gtctacagaa ccaccgtgga ctacacctac ttcagcgagc ccgtgtcctg
     1861 cgaagtgaca aatgccctgg gcagcaccaa cctgtccaga accgtggacg tgtacttcgg
     1921 ccccagaatg accacagagc ctcagagcct gctggtggat ctgggctccg atgccatctt
     1981 tagctgtgcc tggaccggca atcctagcct gaccatcgtg tggatgaagc gcggatctgg
     2041 cgtggtgctg agcaacgaga aaaccctgac actgaagtcc gtgcggcaag aggacgccgg
     2101 caaatacgtg tgtagagccg tggtgcctag agtcggagcc ggcgaaagag aagtgaccct
     2161 gaccgtgaat ggccctccta tcatcagcag cacacagacc cagcatgccc tgcacggcga
     2221 gaagggacag atcaagtgct tcatccgcag cacccctcca cctgacagaa tcgcctggtc
     2281 ttggaaagaa aacgtgctgg aaagcggcac cagcggcaga tacaccgtgg aaaccatcag
     2341 caccgaggaa ggcgtgatca gcacactgac catctccaac atcgtgcggg ccgacttcca
     2401 gaccatctac aattgcacag cctggaacag cttcggcagc gacaccgaga tcatccggct
     2461 gaaagaacag ggcagcgaga tgaagtctgg cgccggactg gaagccgagt ctgttcctat
     2521 ggctggcgcg ccgggcggcg acaagaccca cacctgcccc ccctgcccag ctccagaact
     2581 gctgggaggg ccttccgtgt tcctgttccc ccccaagccc aaggacaccc tgatgatcag
     2641 caggaccccc gaggtgacct gcgtggtggt ggccgtgtcc cacgaggacc cagaggtgaa
     2701 gttcaactgg tacgtggacg gcgtggaggt gcacaacgcc aagaccaagc ccagagagga
     2761 gcagtacaac agcacctaca gggtggtgtc cgtgctgacc gtgctgcacc aggactggct
     2821 gaacggcaaa gaatacaagt gcaaagtctc caacaaggcc ctggctgccc caatcgaaaa
     2881 gacaatcagc aaggccaagg gccagccacg ggagccccag gtgtacaccc tgccccccag
     2941 ccgggaggag atgaccaaga accaggtgtc cctgacctgt ctggtgaagg gcttctaccc
     3001 cagcgatatc gccgtggagt gggagagcaa cggccagccc gagaacaact acaagaccac
     3061 ccccccagtg ctggacagcg acggcagctt cttcctgtac agcaagctga ccgtggacaa
     3121 gtccaggtgg cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa
     3181 ccactacacc cagaagtccc tgagcctgag ccccggcggc agcggtctta atgatatttt
     3241 cgaggctcag aagattgagt ggcatgaaca ccaccaccat caccactaag gccggcctgt
     3301 agtcactggc acaacgcgtt acccggtaag ccaatcgggt atacacggtc gtcatactgc
     3361 agacagggtt cttctacttt gcaagatagt cttgagtagt aaaataaata gatagagaaa
     3421 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3481 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3541 atcacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3601 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3661 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3721 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3841 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3901 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3961 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4021 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     4081 ataatacgac aaggtgagga actaaaccat gaccgagtac aagcccacgg tgcgcctcgc
     4141 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4201 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4261 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4321 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4381 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4441 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4501 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4561 cgagcgcgcc ggggtgcccg ccttcctgga gacatccgcg ccccgcaacc tccccttcta
     4621 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4681 gtgcatgacc cgcaagcccg gtgcctgaca cgtgctacga gatttcgatt ccaccgccgc
     4741 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4801 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4861 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4921 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4981 ctctagctag agcttggcgt aatcatggtc attaccaatg cttaatcagt gaggcaccta
     5041 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5101 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     5161 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5221 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5281 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     5341 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5401 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5461 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5521 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5581 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5641 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5701 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5761 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5821 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     5881 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5941 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     6001 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg ctcatgacca
     6061 aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa
     6121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6241 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6301 gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6841 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6901 ggaaggcgag agtagggaac tgccaggcac tgagagatcc cctcataatt tccccaaagc
     6961 gtaaccatgt gtgaataaat tttgagctag tagggttgca gccacgagta agtcttccct
     7021 tgttattgtg tagccagaat gccgcaaaac ttccatgcct aagcgaactg ttgagagtac
     7081 gtttcgattt ctgactgtgt tagcctggaa gtgcttgtcc caaccttgtt tctgagcatg
     7141 aacgcccgca agccaacatg ttagttgaag catcagggcg attagcagca tgatatcaaa
     7201 acgctctgag ctgctcgttc ggctatggcg taggcctagt ccgtaggcag gacttttcaa
     7261 gtctcggaag gtttcttcaa tctgcattcg cttcgaatag atattaacaa gttgtttggg
     7321 tgttcgaatt tcaacaggta agttagttgc tagaacccat ggctcctttg ccgacgctga
     7381 gtagatttta ggtgacgggt ggtgacaatg agtccgtgtc gagcgctgat tttttcggcc
     7441 tttagagcga gatttataca atagaatttg gcatgagatt ggattgcttt tagtcagcct
     7501 cttatagcct aaagtctttg agtgactaga tgacatatca tgtaagttgc tgataggttt
     7561 ccagttttcc gctcctaggt ctgcatattg tacttttcct cttactcgac ttaaccagta
     7621 ccaacccagc ttctcaacgg atttatacca tggcacttta aagccagcat cactgacaat
     7681 gagcggtgtg gtgttactcg gtagaatgct cgcaaggtcg gctagaaatt ggtcatgagc
     7741 tttctttgaa cattgctctg aaagcgggaa cgctttctca taaagagtaa cagaacgacc
     7801 gtgtagtgcg actgaagctc gcaataccat aagtcgtttt tgctcacgaa tatcagacca
     7861 gtcaacaagt acaatgggca tcgtattgcc cgaacagata aagctagcat gccaacggta
     7921 tacagcgagt cgctctttgt ggaggtgacg attacctaac aatcggtcga ttcgtttgat
     7981 gttatgtttt gttctcgctt tggttggcag gttacggcca agttcggtaa gagtgagagt
     8041 tttacagtca agtaatgcgt ggcaagccaa cgttaagctg ttgagtcgtt ttaagtgtaa
     8101 ttcggggcag aattggtaaa gagagtcgtg taaaatatcg agttcgcaca tcttgttgtc
     8161 tgattattga tttttcgcga aaccatttga tcatatgaca agatgtgtat ccaccttaac
     8221 ttaatgattt ttaccaaaat cattagggga ttcatcagtg ccaggcatca aactaagcag
     8281 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     8341 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     8401 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     8461 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     8521 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     8581 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     8641 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     8701 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     8761 cagtatttat tatcatttag aataaatttt gtgtcgccct taattgtgag cggataacaa
     8821 ttacgagctt catgcacagt ggcgttgact caatattggc cattagccat attattcatt
     8881 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     8941 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgtt
//
