LOCUS       sequence-368748-        6640 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 6640)
  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 6640)
  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-PILRA-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     191..211
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    329..440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     429..446
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(586..603)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(604..648)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1999..2018)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2031..2049)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2032..2049)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2259..2283)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2280..2483)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2309..2329)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2484..2863
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3030..3046
                     /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(3502..3519)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3502..3518)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3617..3634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3788..4376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3868..3887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4462..4481)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4487..4506)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4548..4639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4640..4708,4709..5500)
                     /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(4858..4877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5553..5674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5564..5583)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5618..5637
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5804..6403)
                     /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(5887..5907)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6384..6403
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6422..6469)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6517..6640,1..206))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6531..6640,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6585..6604)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      181 gactttccac accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt
      241 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
      301 aatagaccga gatagggttg agtgttgtga tgcaatttcc tcattttatt aggaaaggac
      361 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
      421 ctggcaacta gaaggcacag tcgaggctga ttttctctat ctatttattt tactactcaa
      481 gactatcttg caaagtagaa gaaccctgtc tgcagtatga cgaccgtgta tacccgattg
      541 gcttaccggg taacgcgttg tgccagtgac tacaggccgg ccttagtggt gatggtggtg
      601 gtgttcatgc cactcaatct tctgagcctc gaaaatatca ttaagaccgc tgccgccggg
      661 gctcaggctc agggacttct gggtgtagtg gttgtgcagg gcctcgtgca tcacgctgca
      721 gctgaacacg ttgccctgct gccacctgga cttgtccacg gtcagcttgc tgtacaggaa
      781 gaagctgccg tcgctgtcca gcactggggg ggtggtcttg tagttgttct cgggctggcc
      841 gttgctctcc cactccacgg cgatatcgct ggggtagaag cccttcacca gacaggtcag
      901 ggacacctgg ttcttggtca tctcctcccg gctggggggc agggtgtaca cctggggctc
      961 ccgtggctgg cccttggcct tgctgattgt cttttcgatt ggggcagcca gggccttgtt
     1021 ggagactttg cacttgtatt ctttgccgtt cagccagtcc tggtgcagca cggtcagcac
     1081 ggacaccacc ctgtaggtgc tgttgtactg ctcctctctg ggcttggtct tggcgttgtg
     1141 cacctccacg ccgtccacgt accagttgaa cttcacctct gggtcctcgt gggacacggc
     1201 caccaccacg caggtcacct cgggggtcct gctgatcatc agggtgtcct tgggcttggg
     1261 ggggaacagg aacacggaag gccctcccag cagttctgga gctgggcagg gggggcaggt
     1321 gtgggtcttg tcgccgcccg gcgcgccagc ggtttccagg ctgatgtgcc agctgtcgct
     1381 tcttctcttg ccctgggtca ctctcaggcc tgttgtggtt gtggtgctgc tcagccgcca
     1441 tgttgtggtc atgctgctag gcctctgtgt tgtggtggtc acggcctggg tgatagacag
     1501 cttggtgccc tcgatgctct gccactgctg tctaccgctg cttctggtgt ccagttccac
     1561 tctgcagaag tacacgctct ggtcctgttt ctgcaggttg ctgattctca ggaagccgga
     1621 cttctggccc tcggtccagt tcaggaacag ccggttcacg tagtccttgt ggatgctagg
     1681 tggtcttgtg ctgtagaagc tctgccggtg gaagtggcct cttctccaag agatccgcac
     1741 atcaggagct gtggcgagtt cccaggggta gtagaagctg aaggggattt ccacgctgcc
     1801 gcccatagag gcagacaggt gcttaggctg tgtcacgcca tacaggtagc tagggccaga
     1861 gccagtagat ccagaaggtt gggcggccgc agcgaacacc aggcagaaga tgtagctcag
     1921 ggcaatgatg gtcttcatgg tggcgctagc aggcgcgttt tcgatagaac cgaggtgcag
     1981 ttggacctgg gagtggacac ctgtggagag aaaggcaaag tggatgttat tctatagtgt
     2041 cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac
     2101 gaagccaagg gggtgggcct atagactcta taggcggtac ttacgtcact cttggcacgg
     2161 ggaatccgcg ttccaatgca ccgttcccgg ccgcggaggc tggatcggtc ccggtgtctt
     2221 ctatggaggt caaaacagcg tggatggcgt ctccaggcga tctgacggtt cactaaacga
     2281 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     2341 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     2401 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     2461 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     2521 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     2581 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     2641 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     2701 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     2761 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     2821 cccgtaattg attactatta ataactagtc aataatcaat gccaacatgg cggtcatatt
     2881 ggacatgagc caatataaat gtacatatta tgatatagat acaacgtatg caatggccaa
     2941 tagccaatat tgatttatgc tatataacca atgaataata tggctaatgg ccaatattga
     3001 gtcaacgcca ctgtgcatga agctcgtaat tgttatccgc tcacaattaa gggcgacaca
     3061 aaatttattc taaatgataa taaatactga taacatctta tagtttgtat tatattttgt
     3121 attatcgttg acatgtataa ttttgatatc aaaaactgat tttcccttta ttattttcga
     3181 gatttatttt cttaattctc tttaacaaac tagaaatatt gtatatacaa aaaatcataa
     3241 ataatagatg aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac gtatcttatt
     3301 taaagtgcgt tgcttttttc tcatttataa ggttaaataa ttctcatata tcaagcaaag
     3361 tgacaggcgc ccttaaatat tctgacaaat gctctttccc taaactcccc ccataaaaaa
     3421 acccgccgaa gcgggttttt acgttatttg cggattaacg attactcgtt atcagaaccg
     3481 cccagggggc ccgagcttaa gactggccgt cgttttacaa cacagaaaga gtttgtagaa
     3541 acgcaaaaag gccatccgtc aggggccttc tgcttagttt gatgcctggc agttccctac
     3601 tctcgccttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4201 ggtgggctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc gtaactcacg
     4441 ttaagggatt ttggtcatga gcaaggggtg ttatgagcca tattcaggta taaatgggct
     4501 cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt gtttattttt
     4561 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4621 atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta ttcccttttt
     4681 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4741 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4801 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4861 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4921 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4981 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5041 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5101 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5161 cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac tattaactgg
     5221 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5281 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatccgg
     5341 agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg gtaagccctc
     5401 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5461 gatcgctgag ataggtgcct cactgattaa gcattggtaa tgaccatgat tacgccaagc
     5521 tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac
     5581 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     5641 atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga
     5701 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     5761 tttcatagaa ggcggcggtg gaatcgaaat ctcgtagcac gtgtcaggca ccgggcttgc
     5821 gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc
     5881 cgagccgctc gtagaagggg aggttgcggg gcgcggatgt ctccaggaag gcgggcaccc
     5941 cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga cccttgccct
     6001 ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg
     6061 gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca
     6121 tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc gtggtccaga
     6181 ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg
     6241 tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt
     6301 ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt
     6361 cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catggtttag ttcctcacct
     6421 tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca cgtgctgatc
     6481 agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa
     6541 aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata
     6601 aataaaaaaa attagtcagc catggggcgg agaatgggcg
//
