LOCUS       sequence-368813-        6457 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 6457)
  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 6457)
  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-NCR3-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(145..192)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(240..569)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      254..389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(308..327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     554..574
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    692..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     792..809
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(949..966)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(967..1011)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2179..2198)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2211..2229)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2212..2229)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2439..2463)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2460..2663)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2489..2509)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2664..3043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3210..3226
                     /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(3682..3699)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3682..3698)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3797..3814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3968..4556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4048..4067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4642..4661)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4667..4686)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4728..4819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4820..4888,4889..5680)
                     /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(5038..5057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5733..5854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5744..5763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5798..5817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5984..6457,1..126))
                     /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(6067..6087)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
       61 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
      121 ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
      181 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
      241 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
      301 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      361 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      421 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      481 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      541 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
      601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      661 aagaatagac cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag
      721 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      781 tggctggcaa ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact
      841 caagactatc ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga
      901 ttggcttacc gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg
      961 gtggtgttca tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc
     1021 ggggctcagg ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct
     1081 gcagctgaac acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag
     1141 gaagaagctg ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg
     1201 gccgttgctc tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt
     1261 cagggacacc tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg
     1321 ctcccgtggc tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt
     1381 gttggagact ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag
     1441 cacggacacc accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt
     1501 gtgcacctcc acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac
     1561 ggccaccacc acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt
     1621 gggggggaac aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca
     1681 ggtgtgggtc ttgtcgccgc ccggcgcgcc tccgagctga gggtgctctt tttccaccac
     1741 cagtcttgtt ccattgcctg tgccaacgcc gaggccgagc acttccactc tacacacgta
     1801 gatgctggcg tcgtgtcccc gaacatctct gatgtgcagc tcggcctggt gatcatgcag
     1861 aaatctgctg ctggccagag gggccagcct tcctctaaac tctggtgtgc cgttccgcac
     1921 ttctttgccg ggcaccacct catctctaaa ccaggtcacg ctgccgatgg ccagtctgcc
     1981 ctgagaggca ttaaagctac aaggcaggaa ggcgctgctg ccttccaggg ttctaatctc
     2041 aggaggctgg gaaacccaca aggcggccgc agcgaacacc aggcagaaga tgtagctcag
     2101 ggcaatgatg gtcttcatgg tggcgctagc aggcgcgttt tcgatagaac cgaggtgcag
     2161 ttggacctgg gagtggacac ctgtggagag aaaggcaaag tggatgttat tctatagtgt
     2221 cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac
     2281 gaagccaagg gggtgggcct atagactcta taggcggtac ttacgtcact cttggcacgg
     2341 ggaatccgcg ttccaatgca ccgttcccgg ccgcggaggc tggatcggtc ccggtgtctt
     2401 ctatggaggt caaaacagcg tggatggcgt ctccaggcga tctgacggtt cactaaacga
     2461 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     2521 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     2581 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     2641 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     2701 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     2761 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     2821 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     2881 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     2941 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     3001 cccgtaattg attactatta ataactagtc aataatcaat gccaacatgg cggtcatatt
     3061 ggacatgagc caatataaat gtacatatta tgatatagat acaacgtatg caatggccaa
     3121 tagccaatat tgatttatgc tatataacca atgaataata tggctaatgg ccaatattga
     3181 gtcaacgcca ctgtgcatga agctcgtaat tgttatccgc tcacaattaa gggcgacaca
     3241 aaatttattc taaatgataa taaatactga taacatctta tagtttgtat tatattttgt
     3301 attatcgttg acatgtataa ttttgatatc aaaaactgat tttcccttta ttattttcga
     3361 gatttatttt cttaattctc tttaacaaac tagaaatatt gtatatacaa aaaatcataa
     3421 ataatagatg aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac gtatcttatt
     3481 taaagtgcgt tgcttttttc tcatttataa ggttaaataa ttctcatata tcaagcaaag
     3541 tgacaggcgc ccttaaatat tctgacaaat gctctttccc taaactcccc ccataaaaaa
     3601 acccgccgaa gcgggttttt acgttatttg cggattaacg attactcgtt atcagaaccg
     3661 cccagggggc ccgagcttaa gactggccgt cgttttacaa cacagaaaga gtttgtagaa
     3721 acgcaaaaag gccatccgtc aggggccttc tgcttagttt gatgcctggc agttccctac
     3781 tctcgccttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4381 ggtgggctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc gtaactcacg
     4621 ttaagggatt ttggtcatga gcaaggggtg ttatgagcca tattcaggta taaatgggct
     4681 cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt gtttattttt
     4741 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4801 atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta ttcccttttt
     4861 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4921 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4981 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5041 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5101 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5161 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5221 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5281 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5341 cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac tattaactgg
     5401 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5461 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatccgg
     5521 agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg gtaagccctc
     5581 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5641 gatcgctgag ataggtgcct cactgattaa gcattggtaa tgaccatgat tacgccaagc
     5701 tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac
     5761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     5821 atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga
     5881 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     5941 tttcatagaa ggcggcggtg gaatcgaaat ctcgtagcac gtgtcaggca ccgggcttgc
     6001 gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc
     6061 cgagccgctc gtagaagggg aggttgcggg gcgcggatgt ctccaggaag gcgggcaccc
     6121 cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga cccttgccct
     6181 ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg
     6241 gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca
     6301 tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc gtggtccaga
     6361 ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg
     6421 tgaggaagag ttcttgcagc tcggtgaccc gctcgat
//
