LOCUS       sequence-368736-        6616 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 6616)
  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 6616)
  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-LSR-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..185
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    303..414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     403..420
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(560..577)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(578..622)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1949..1968)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1981..1999)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1982..1999)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2209..2233)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2230..2433)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2259..2279)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2434..2813
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2980..2996
                     /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)."
     primer_bind     complement(3452..3469)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3452..3468)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3567..3584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3738..4326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3818..3837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4412..4431)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4437..4456)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4498..4589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4590..4658,4659..5450)
                     /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     complement(4808..4827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5503..5624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5514..5533)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5568..5587
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5754..6353)
                     /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(5837..5857)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6334..6353
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6372..6419)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6467..6616,1..180))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6481..6616
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6535..6554)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
       61 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
      121 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
      181 attccacaga attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
      241 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
      301 gtgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca
      361 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg
      421 ctgattttct ctatctattt attttactac tcaagactat cttgcaaagt agaagaaccc
      481 tgtctgcagt atgacgaccg tgtatacccg attggcttac cgggtaacgc gttgtgccag
      541 tgactacagg ccggccttag tggtgatggt ggtggtgttc atgccactca atcttctgag
      601 cctcgaaaat atcattaaga ccgctgccgc cggggctcag gctcagggac ttctgggtgt
      661 agtggttgtg cagggcctcg tgcatcacgc tgcagctgaa cacgttgccc tgctgccacc
      721 tggacttgtc cacggtcagc ttgctgtaca ggaagaagct gccgtcgctg tccagcactg
      781 ggggggtggt cttgtagttg ttctcgggct ggccgttgct ctcccactcc acggcgatat
      841 cgctggggta gaagcccttc accagacagg tcagggacac ctggttcttg gtcatctcct
      901 cccggctggg gggcagggtg tacacctggg gctcccgtgg ctggcccttg gccttgctga
      961 ttgtcttttc gattggggca gccagggcct tgttggagac tttgcacttg tattctttgc
     1021 cgttcagcca gtcctggtgc agcacggtca gcacggacac caccctgtag gtgctgttgt
     1081 actgctcctc tctgggcttg gtcttggcgt tgtgcacctc cacgccgtcc acgtaccagt
     1141 tgaacttcac ctctgggtcc tcgtgggaca cggccaccac cacgcaggtc acctcggggg
     1201 tcctgctgat catcagggtg tccttgggct tgggggggaa caggaacacg gaaggccctc
     1261 ccagcagttc tggagctggg cagggggggc aggtgtgggt cttgtcgccg cccggcgcgc
     1321 catcctcgat aggtccggcc tgaaatccag gcagcagctc agcaactccg cttgttctgc
     1381 ccagcacgat cagctcggca taggcttcat tgttgccctg caggtcctgg gcagacacca
     1441 cagaacagta gtacacgccg ctgtcgcccc aagctgtctg atcaaaggtc aggtcggcgt
     1501 tgccggtgat ggtgattctt ctgccctggt agtagtcgcc cagtgtcacg gcatttccct
     1561 gctttgtggc caccactctg actgtccgca cgctgtcctg gcattccaca taaggattgt
     1621 agccagggtt gccagcggcc agctgagcat tcagctgatt gtccacggag gcgggagaaa
     1681 aggcatcggc gattctgtcc cggcagaagc tcttgtactt ccagatcacg atgggctgtg
     1741 taggtgtgct ggtcatctgg taggtgcagg gcagtgtcac aggctggaac aggatcacga
     1801 cgtggtaagg attggacacg gtcacttgaa tggcggccgc agcgaacacc aggcagaaga
     1861 tgtagctcag ggcaatgatg gtcttcatgg tggcgctagc aggcgcgttt tcgatagaac
     1921 cgaggtgcag ttggacctgg gagtggacac ctgtggagag aaaggcaaag tggatgttat
     1981 tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc
     2041 gcgttctaac gaagccaagg gggtgggcct atagactcta taggcggtac ttacgtcact
     2101 cttggcacgg ggaatccgcg ttccaatgca ccgttcccgg ccgcggaggc tggatcggtc
     2161 ccggtgtctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga tctgacggtt
     2221 cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     2281 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2341 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     2401 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     2461 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     2521 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     2581 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     2641 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     2701 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     2761 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gccaacatgg
     2821 cggtcatatt ggacatgagc caatataaat gtacatatta tgatatagat acaacgtatg
     2881 caatggccaa tagccaatat tgatttatgc tatataacca atgaataata tggctaatgg
     2941 ccaatattga gtcaacgcca ctgtgcatga agctcgtaat tgttatccgc tcacaattaa
     3001 gggcgacaca aaatttattc taaatgataa taaatactga taacatctta tagtttgtat
     3061 tatattttgt attatcgttg acatgtataa ttttgatatc aaaaactgat tttcccttta
     3121 ttattttcga gatttatttt cttaattctc tttaacaaac tagaaatatt gtatatacaa
     3181 aaaatcataa ataatagatg aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac
     3241 gtatcttatt taaagtgcgt tgcttttttc tcatttataa ggttaaataa ttctcatata
     3301 tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat gctctttccc taaactcccc
     3361 ccataaaaaa acccgccgaa gcgggttttt acgttatttg cggattaacg attactcgtt
     3421 atcagaaccg cccagggggc ccgagcttaa gactggccgt cgttttacaa cacagaaaga
     3481 gtttgtagaa acgcaaaaag gccatccgtc aggggccttc tgcttagttt gatgcctggc
     3541 agttccctac tctcgccttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3601 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3661 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3721 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3781 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3841 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3901 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3961 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4021 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4081 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4141 ttcttgaagt ggtgggctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4201 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4261 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4321 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc
     4381 gtaactcacg ttaagggatt ttggtcatga gcaaggggtg ttatgagcca tattcaggta
     4441 taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt
     4501 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4561 tgcttcaata atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta
     4621 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4681 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4741 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4801 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4861 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4921 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4981 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5041 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5101 taccaaacga cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac
     5161 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5221 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5281 ataaatccgg agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg
     5341 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5401 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa tgaccatgat
     5461 tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt
     5521 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     5581 ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac
     5641 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     5701 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtagcac gtgtcaggca
     5761 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     5821 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggatgt ctccaggaag
     5881 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     5941 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     6001 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     6061 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     6121 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     6181 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga
     6241 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
     6301 ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catggtttag
     6361 ttcctcacct tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca
     6421 cgtgctgatc agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta
     6481 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg
     6541 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     6601 gagttagggg cgggat
//
