LOCUS       sequence-368534-        6598 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 6598)
  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 6598)
  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-TNFRSF14-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     242..262
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    380..491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     480..497
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(637..654)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(655..699)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2008..2027)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2040..2058)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2041..2058)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2268..2292)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2289..2492)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2318..2338)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2493..2872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3039..3055
                     /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(3511..3528)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3511..3527)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3626..3643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3797..4385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3877..3896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4471..4490)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4496..4515)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4557..4648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4649..4717,4718..5509)
                     /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(4867..4886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5562..5683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5573..5592)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5627..5646
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5813..6412)
                     /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(5896..5916)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6393..6412
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6431..6478)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6526..6598,1..257))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6540..6598,1..77)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(6594..6598,1..15))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
       61 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
      121 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
      181 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      241 caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt taaatcagct
      301 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
      361 agatagggtt gagtgttgtg atgcaatttc ctcattttat taggaaagga cagtgggagt
      421 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
      481 agaaggcaca gtcgaggctg attttctcta tctatttatt ttactactca agactatctt
      541 gcaaagtaga agaaccctgt ctgcagtatg acgaccgtgt atacccgatt ggcttaccgg
      601 gtaacgcgtt gtgccagtga ctacaggccg gccttagtgg tgatggtggt ggtgttcatg
      661 ccactcaatc ttctgagcct cgaaaatatc attaagaccg ctgccgccgg ggctcaggct
      721 cagggacttc tgggtgtagt ggttgtgcag ggcctcgtgc atcacgctgc agctgaacac
      781 gttgccctgc tgccacctgg acttgtccac ggtcagcttg ctgtacagga agaagctgcc
      841 gtcgctgtcc agcactgggg gggtggtctt gtagttgttc tcgggctggc cgttgctctc
      901 ccactccacg gcgatatcgc tggggtagaa gcccttcacc agacaggtca gggacacctg
      961 gttcttggtc atctcctccc ggctgggggg cagggtgtac acctggggct cccgtggctg
     1021 gcccttggcc ttgctgattg tcttttcgat tggggcagcc agggccttgt tggagacttt
     1081 gcacttgtat tctttgccgt tcagccagtc ctggtgcagc acggtcagca cggacaccac
     1141 cctgtaggtg ctgttgtact gctcctctct gggcttggtc ttggcgttgt gcacctccac
     1201 gccgtccacg taccagttga acttcacctc tgggtcctcg tgggacacgg ccaccaccac
     1261 gcaggtcacc tcgggggtcc tgctgatcat cagggtgtcc ttgggcttgg gggggaacag
     1321 gaacacggaa ggccctccca gcagttctgg agctgggcag ggggggcagg tgtgggtctt
     1381 gtcgccgccc ggcgcgccaa cccaatggct gctgcttgtt ccggcgccag cctttgtaac
     1441 cagccagcta cacttggtct ggtgctggca ctcttccagt gtgccgttgg ggctgaatgt
     1501 gcctggaggg caattttgac acagggtatc ctggctctcg gtgccgcctt tctgcactct
     1561 ctgtccaggg ctagatgtgg cataggctct acaggcggca cagtgatcgc catcctgcac
     1621 gatacagaag tggccagggc tacagccaca cacggcattc tcggttctgc tgcagtttct
     1681 gctggctctc agtcccatgg caggatcaca catctggcac tgcaggcact tgctcaggcc
     1741 gttcaggtgg gcgatatatg tgccaggagg acaaggttca cacacggtgc cggtcagttc
     1801 tccgcaggcc tctttcactc tgtagccggg agaacactta gggcagcact cgctgccgac
     1861 aggatactca tcctctttgc agctaggcaa ggcggccgca gcgaacacca ggcagaagat
     1921 gtagctcagg gcaatgatgg tcttcatggt ggcgctagca ggcgcgtttt cgatagaacc
     1981 gaggtgcagt tggacctggg agtggacacc tgtggagaga aaggcaaagt ggatgttatt
     2041 ctatagtgtc acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg
     2101 cgttctaacg aagccaaggg ggtgggccta tagactctat aggcggtact tacgtcactc
     2161 ttggcacggg gaatccgcgt tccaatgcac cgttcccggc cgcggaggct ggatcggtcc
     2221 cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc
     2281 actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     2341 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     2401 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     2461 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     2521 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     2581 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     2641 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     2701 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     2761 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     2821 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg ccaacatggc
     2881 ggtcatattg gacatgagcc aatataaatg tacatattat gatatagata caacgtatgc
     2941 aatggccaat agccaatatt gatttatgct atataaccaa tgaataatat ggctaatggc
     3001 caatattgag tcaacgccac tgtgcatgaa gctcgtaatt gttatccgct cacaattaag
     3061 ggcgacacaa aatttattct aaatgataat aaatactgat aacatcttat agtttgtatt
     3121 atattttgta ttatcgttga catgtataat tttgatatca aaaactgatt ttccctttat
     3181 tattttcgag atttattttc ttaattctct ttaacaaact agaaatattg tatatacaaa
     3241 aaatcataaa taatagatga atagtttaat tataggtgtt catcaatcga aaaagcaacg
     3301 tatcttattt aaagtgcgtt gcttttttct catttataag gttaaataat tctcatatat
     3361 caagcaaagt gacaggcgcc cttaaatatt ctgacaaatg ctctttccct aaactccccc
     3421 cataaaaaaa cccgccgaag cgggttttta cgttatttgc ggattaacga ttactcgtta
     3481 tcagaaccgc ccagggggcc cgagcttaag actggccgtc gttttacaac acagaaagag
     3541 tttgtagaaa cgcaaaaagg ccatccgtca ggggccttct gcttagtttg atgcctggca
     3601 gttccctact ctcgccttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3661 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3721 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3781 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3841 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3901 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3961 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4021 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4081 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4141 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4201 tcttgaagtg gtgggctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4261 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4321 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4381 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg
     4441 taactcacgt taagggattt tggtcatgag caaggggtgt tatgagccat attcaggtat
     4501 aaatgggctc gcgataatgt tcagaattgg ttaattggtt gtaacactga cccctatttg
     4561 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4621 gcttcaataa tattgaaaaa ggaagaatat gagtattcaa catttccgtg tcgcccttat
     4681 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4741 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4801 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4861 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4921 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4981 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5041 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5101 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5161 accaaacgac gagcgtgaca ccacgatgcc tgtagcgatg gcaacaacgt tgcgcaaact
     5221 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5281 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5341 taaatccgga gccggtgagc gtggttctcg cggtatcatc gcagcgctgg ggccagatgg
     5401 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5461 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat gaccatgatt
     5521 acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt
     5581 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     5641 tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact
     5701 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     5761 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg tgtcaggcac
     5821 cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg tcggcggtga
     5881 cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggatgtc tccaggaagg
     5941 cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg ctgcccagac
     6001 ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg ggctccttgg
     6061 gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg gaaccgctca
     6121 actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg ctctccggcg
     6181 tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc
     6241 cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg cggtccggat
     6301 cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc
     6361 tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc atggtttagt
     6421 tcctcacctt gtcgtattat actatgccga tatactatgc cgatgattaa ttgtcaacac
     6481 gtgctgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     6541 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcg
//
