LOCUS       sequence-368530-        6595 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 6595)
  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 6595)
  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-TNFRSF9-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    386..497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     486..503
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(643..660)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(661..705)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2011..2030)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2043..2061)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2044..2061)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2271..2295)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2292..2495)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2321..2341)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2496..2875
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3042..3058
                     /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(3514..3531)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3514..3530)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3629..3646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3800..4388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3880..3899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4474..4493)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4499..4518)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4560..4651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4652..4720,4721..5512)
                     /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(4870..4889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5565..5686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5576..5595)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5630..5649
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5816..6415)
                     /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(5899..5919)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6396..6415
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6434..6481)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6529..6595,1..263))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6543..6595,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
      301 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      361 agaccgagat agggttgagt gttgtgatgc aatttcctca ttttattagg aaaggacagt
      421 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
      481 gcaactagaa ggcacagtcg aggctgattt tctctatcta tttattttac tactcaagac
      541 tatcttgcaa agtagaagaa ccctgtctgc agtatgacga ccgtgtatac ccgattggct
      601 taccgggtaa cgcgttgtgc cagtgactac aggccggcct tagtggtgat ggtggtggtg
      661 ttcatgccac tcaatcttct gagcctcgaa aatatcatta agaccgctgc cgccggggct
      721 caggctcagg gacttctggg tgtagtggtt gtgcagggcc tcgtgcatca cgctgcagct
      781 gaacacgttg ccctgctgcc acctggactt gtccacggtc agcttgctgt acaggaagaa
      841 gctgccgtcg ctgtccagca ctgggggggt ggtcttgtag ttgttctcgg gctggccgtt
      901 gctctcccac tccacggcga tatcgctggg gtagaagccc ttcaccagac aggtcaggga
      961 cacctggttc ttggtcatct cctcccggct ggggggcagg gtgtacacct ggggctcccg
     1021 tggctggccc ttggccttgc tgattgtctt ttcgattggg gcagccaggg ccttgttgga
     1081 gactttgcac ttgtattctt tgccgttcag ccagtcctgg tgcagcacgg tcagcacgga
     1141 caccaccctg taggtgctgt tgtactgctc ctctctgggc ttggtcttgg cgttgtgcac
     1201 ctccacgccg tccacgtacc agttgaactt cacctctggg tcctcgtggg acacggccac
     1261 caccacgcag gtcacctcgg gggtcctgct gatcatcagg gtgtccttgg gcttgggggg
     1321 gaacaggaac acggaaggcc ctcccagcag ttctggagct gggcaggggg ggcaggtgtg
     1381 ggtcttgtcg ccgcccggcg cgccttgagg agaatggcca ggctctctgg ctggagcagg
     1441 aggtgtaacg ctagaagcgc caggagacag atcagcagga gatgggccac acacgacatc
     1501 cctttctttg gtgccgttga ccagcacgct cttgccatcc aggctgcaat tggtccaagg
     1561 ccggcagatg cctctcttct ggtcgttaaa ggtgccgaag cagcaatcct tgcagccctt
     1621 cttggtcagc tcttggccct gcttgcaatc ctgctcgcac atagaacagc cggctccgag
     1681 acagtgaaag ccaggggtgc aatcacactc ggcattgctg gtgctgctgc actcttttct
     1741 ggtccggaac acgcccttgc actgtctgca gatgtcgcag gttctctgtc cgccggcaga
     1801 gctaaagctg ttaggaggac aggggctgca gatctggttt ctgttgttgt cgcagaaggt
     1861 gccggcagga caattagaac aagggtcctg caaggcggcc gcagcgaaca ccaggcagaa
     1921 gatgtagctc agggcaatga tggtcttcat ggtggcgcta gcaggcgcgt tttcgataga
     1981 accgaggtgc agttggacct gggagtggac acctgtggag agaaaggcaa agtggatgtt
     2041 attctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag
     2101 ccgcgttcta acgaagccaa gggggtgggc ctatagactc tataggcggt acttacgtca
     2161 ctcttggcac ggggaatccg cgttccaatg caccgttccc ggccgcggag gctggatcgg
     2221 tcccggtgtc ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg
     2281 ttcactaaac gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     2341 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     2401 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     2461 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     2521 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     2581 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     2641 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     2701 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     2761 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     2821 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgccaacat
     2881 ggcggtcata ttggacatga gccaatataa atgtacatat tatgatatag atacaacgta
     2941 tgcaatggcc aatagccaat attgatttat gctatataac caatgaataa tatggctaat
     3001 ggccaatatt gagtcaacgc cactgtgcat gaagctcgta attgttatcc gctcacaatt
     3061 aagggcgaca caaaatttat tctaaatgat aataaatact gataacatct tatagtttgt
     3121 attatatttt gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt
     3181 tattattttc gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac
     3241 aaaaaatcat aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca
     3301 acgtatctta tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata
     3361 tatcaagcaa agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc
     3421 ccccataaaa aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg
     3481 ttatcagaac cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa
     3541 gagtttgtag aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg
     3601 gcagttccct actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3661 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3721 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3781 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3841 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3901 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3961 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4021 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4141 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4201 agttcttgaa gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg
     4261 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4321 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4381 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc
     4441 gcgtaactca cgttaaggga ttttggtcat gagcaagggg tgttatgagc catattcagg
     4501 tataaatggg ctcgcgataa tgttcagaat tggttaattg gttgtaacac tgacccctat
     4561 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4621 aatgcttcaa taatattgaa aaaggaagaa tatgagtatt caacatttcc gtgtcgccct
     4681 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4741 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4801 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4861 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4921 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4981 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5041 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5101 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5161 cataccaaac gacgagcgtg acaccacgat gcctgtagcg atggcaacaa cgttgcgcaa
     5221 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5281 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5341 tgataaatcc ggagccggtg agcgtggttc tcgcggtatc atcgcagcgc tggggccaga
     5401 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5461 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aatgaccatg
     5521 attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga
     5581 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     5641 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga
     5701 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     5761 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtagc acgtgtcagg
     5821 caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg
     5881 tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggat gtctccagga
     5941 aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca
     6001 gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct
     6061 tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc
     6121 tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg
     6181 gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga
     6241 gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg
     6301 gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
     6361 ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggttt
     6421 agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
     6481 cacgtgctga tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc
     6541 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcg
//
