LOCUS       sequence-368741-        6622 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 6622)
  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 6622)
  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-TNFSF8-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     239..259
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    377..488
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     477..494
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(634..651)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(652..696)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2029..2048)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2061..2079)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2062..2079)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2289..2313)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2310..2513)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2339..2359)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2514..2893
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3060..3076
                     /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(3532..3549)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3532..3548)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3647..3664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3818..4406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3898..3917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4492..4511)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4517..4536)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4578..4669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4670..4738,4739..5530)
                     /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(4888..4907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5583..5704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5594..5613)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5648..5667
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5834..6433)
                     /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(5917..5937)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6414..6433
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6452..6499)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6547..6622,1..254))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6561..6622,1..74)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(6615..6622,1..12))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
       61 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
      121 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
      181 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      241 cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa atcagctcat
      301 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      361 tagggttgag tgttgtgatg caatttcctc attttattag gaaaggacag tgggagtggc
      421 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
      481 aggcacagtc gaggctgatt ttctctatct atttatttta ctactcaaga ctatcttgca
      541 aagtagaaga accctgtctg cagtatgacg accgtgtata cccgattggc ttaccgggta
      601 acgcgttgtg ccagtgacta caggccggcc ttagtggtga tggtggtggt gttcatgcca
      661 ctcaatcttc tgagcctcga aaatatcatt aagaccgctg ccgccggggc tcaggctcag
      721 ggacttctgg gtgtagtggt tgtgcagggc ctcgtgcatc acgctgcagc tgaacacgtt
      781 gccctgctgc cacctggact tgtccacggt cagcttgctg tacaggaaga agctgccgtc
      841 gctgtccagc actggggggg tggtcttgta gttgttctcg ggctggccgt tgctctccca
      901 ctccacggcg atatcgctgg ggtagaagcc cttcaccaga caggtcaggg acacctggtt
      961 cttggtcatc tcctcccggc tggggggcag ggtgtacacc tggggctccc gtggctggcc
     1021 cttggccttg ctgattgtct tttcgattgg ggcagccagg gccttgttgg agactttgca
     1081 cttgtattct ttgccgttca gccagtcctg gtgcagcacg gtcagcacgg acaccaccct
     1141 gtaggtgctg ttgtactgct cctctctggg cttggtcttg gcgttgtgca cctccacgcc
     1201 gtccacgtac cagttgaact tcacctctgg gtcctcgtgg gacacggcca ccaccacgca
     1261 ggtcacctcg ggggtcctgc tgatcatcag ggtgtccttg ggcttggggg ggaacaggaa
     1321 cacggaaggc cctcccagca gttctggagc tgggcagggg gggcaggtgt gggtcttgtc
     1381 gccgcccggc gcgccatcgg agttgctgta caggaagatg ctcagcacgt tttccagtgg
     1441 aaaggtgctg gtgtcgatgt actggaaggt gtccacgttc acgctgatgg tggtgttcac
     1501 ttgcaggtag tccagcagga actggctcag gttctggtac acgtgcttgg tctgcattcc
     1561 gctctcgcac actgtgacca gggcctgctt cttgatgtgc ttgttgatca gcagttccag
     1621 cttcaggtcc acgctattgt tggggcactg caccaggaac tgcagctggc agataatgaa
     1681 gtacaggccg gggaactgaa tcacgaggtt gccatcctgg tagcgcacgc catgcaggat
     1741 gccgtccttg ttccagctca gcttggtctt gttcaggtgc ttggccacct ggaggtaggc
     1801 ccaggacttc ttgaaagggg ctctcttcag aatgcacagc agatcctcgc tgcaattgcc
     1861 gcctttcagg ggcacgttgt cggggctgtt agggatagaa tcggttctct gggcggccgc
     1921 agcgaacacc aggcagaaga tgtagctcag ggcaatgatg gtcttcatgg tggcgctagc
     1981 aggcgcgttt tcgatagaac cgaggtgcag ttggacctgg gagtggacac ctgtggagag
     2041 aaaggcaaag tggatgttat tctatagtgt cacctaaatc gtatgtgtat gatacataag
     2101 gttatgtatt aattgtagcc gcgttctaac gaagccaagg gggtgggcct atagactcta
     2161 taggcggtac ttacgtcact cttggcacgg ggaatccgcg ttccaatgca ccgttcccgg
     2221 ccgcggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg tggatggcgt
     2281 ctccaggcga tctgacggtt cactaaacga gctctgctta tatagacctc ccaccgtaca
     2341 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     2401 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     2461 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     2521 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     2581 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     2641 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     2701 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     2761 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     2821 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     2881 aataatcaat gccaacatgg cggtcatatt ggacatgagc caatataaat gtacatatta
     2941 tgatatagat acaacgtatg caatggccaa tagccaatat tgatttatgc tatataacca
     3001 atgaataata tggctaatgg ccaatattga gtcaacgcca ctgtgcatga agctcgtaat
     3061 tgttatccgc tcacaattaa gggcgacaca aaatttattc taaatgataa taaatactga
     3121 taacatctta tagtttgtat tatattttgt attatcgttg acatgtataa ttttgatatc
     3181 aaaaactgat tttcccttta ttattttcga gatttatttt cttaattctc tttaacaaac
     3241 tagaaatatt gtatatacaa aaaatcataa ataatagatg aatagtttaa ttataggtgt
     3301 tcatcaatcg aaaaagcaac gtatcttatt taaagtgcgt tgcttttttc tcatttataa
     3361 ggttaaataa ttctcatata tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat
     3421 gctctttccc taaactcccc ccataaaaaa acccgccgaa gcgggttttt acgttatttg
     3481 cggattaacg attactcgtt atcagaaccg cccagggggc ccgagcttaa gactggccgt
     3541 cgttttacaa cacagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggggccttc
     3601 tgcttagttt gatgcctggc agttccctac tctcgccttc cgcttcctcg ctcactgact
     3661 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3721 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4201 atgtaggcgg tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa
     4261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4381 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4441 ctcagtggaa cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gcaaggggtg
     4501 ttatgagcca tattcaggta taaatgggct cgcgataatg ttcagaattg gttaattggt
     4561 tgtaacactg acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagaata tgagtattca
     4681 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4741 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4801 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4861 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4921 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4981 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5041 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5101 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5161 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcgat
     5221 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5281 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5341 ggctggctgg tttattgctg ataaatccgg agccggtgag cgtggttctc gcggtatcat
     5401 cgcagcgctg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5461 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5521 gcattggtaa tgaccatgat tacgccaagc tctagctaga ggtcgacggt atacagacat
     5581 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     5641 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     5701 agttggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     5761 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     5821 ctcgtagcac gtgtcaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg
     5881 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
     5941 gcgcggatgt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
     6001 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca
     6061 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
     6121 cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc
     6181 ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc
     6241 acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc
     6301 gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg
     6361 cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct
     6421 tgtactcggt catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg
     6481 ccgatgatta attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg
     6541 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     6601 agaggcagag gcggcctcgg cc
//
