LOCUS       sequence-371819-        6340 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 6340)
  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 6340)
  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-TNFRSF13C-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        88..135
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             154..753
                     /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(154..173)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     650..670
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    883..1004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(920..939)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     974..993
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1057..1848,1849..1917))
                     /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     1680..1699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1918..2009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2051..2070
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2076..2095
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2181..2769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2670..2689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2923..2940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3038..3055
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3039..3055
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3511..3527
                     /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)."
     enhancer        3694..4073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4074..4277
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4228..4248
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4274..4298
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4508..4526
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4508..4525
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4539..4558
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5609..5653
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5654..5671
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5811..5828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5817..5928
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6046..6066)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6051..6340,1..40)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6231..6340,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6293..6312
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
       61 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
      121 atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc cacggtgcgc
      181 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
      241 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
      301 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
      361 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
      421 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
      481 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
      541 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
      601 gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg caacctcccc
      661 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
      721 acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt cgattccacc
      781 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
      841 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
      901 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
      961 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     1021 tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa tcagtgaggc
     1081 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1141 gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga taccgcgaga
     1201 accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa gggccgagcg
     1261 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1321 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg ctacaggcat
     1381 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1441 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1501 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1561 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     1621 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1681 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1741 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1801 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1861 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatatt
     1921 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1981 atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca attaaccaat
     2041 tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc ccttgctcat
     2101 gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc agaccccgta
     2161 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2221 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2281 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2341 ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2401 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2461 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2521 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2581 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2641 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2701 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2761 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2821 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2881 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2941 gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag gcccctgacg
     3001 gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg ccagtcttaa
     3061 gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa taacgtaaaa
     3121 acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt gtcagaatat
     3181 ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat aaatgagaaa
     3241 aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac ctataattaa
     3301 actattcatc tattatttat gattttttgt atatacaata tttctagttt gttaaagaga
     3361 attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata tcaaaattat
     3421 acatgtcaac gataatacaa aatataatac aaactataag atgttatcag tatttattat
     3481 catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta cgagcttcat
     3541 gcacagtggc gttgactcaa tattggccat tagccatatt attcattggt tatatagcat
     3601 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     3661 atattggctc atgtccaata tgaccgccat gttggcattg attattgact agttattaat
     3721 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     3781 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     3841 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     3901 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     3961 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4021 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     4081 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4141 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4201 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4261 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
     4321 gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggccgg gaacggtgca
     4381 ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc
     4441 ccaccccctt ggcttcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     4501 acatacgatt taggtgacac tatagaataa catccacttt gcctttctct ccacaggtgt
     4561 ccactcccag gtccaactgc acctcggttc tatcgaaaac gcgcctgcta gcgccaccat
     4621 gaagaccatc attgccctga gctacatctt ctgcctggtg ttcgctgcgg ccgccatgag
     4681 aagaggccct agatctctga gaggcagaga tgcccctgct cctacacctt gtgtgcctgc
     4741 cgagtgcttt gatctgctcg tcagacactg tgtggcctgc ggactgctga gaacccctag
     4801 acctaaacct gccggcgctt ctagccctgc tccaagaaca gctctgcagc ctcaagagtc
     4861 tgttggagct ggtgctggcg aagctgcttt gcctctgcct ggacttcttg gcgcgccggg
     4921 cggcgacaag acccacacct gccccccctg cccagctcca gaactgctgg gagggccttc
     4981 cgtgttcctg ttccccccca agcccaagga caccctgatg atcagcagga cccccgaggt
     5041 gacctgcgtg gtggtggccg tgtcccacga ggacccagag gtgaagttca actggtacgt
     5101 ggacggcgtg gaggtgcaca acgccaagac caagcccaga gaggagcagt acaacagcac
     5161 ctacagggtg gtgtccgtgc tgaccgtgct gcaccaggac tggctgaacg gcaaagaata
     5221 caagtgcaaa gtctccaaca aggccctggc tgccccaatc gaaaagacaa tcagcaaggc
     5281 caagggccag ccacgggagc cccaggtgta caccctgccc cccagccggg aggagatgac
     5341 caagaaccag gtgtccctga cctgtctggt gaagggcttc taccccagcg atatcgccgt
     5401 ggagtgggag agcaacggcc agcccgagaa caactacaag accacccccc cagtgctgga
     5461 cagcgacggc agcttcttcc tgtacagcaa gctgaccgtg gacaagtcca ggtggcagca
     5521 gggcaacgtg ttcagctgca gcgtgatgca cgaggccctg cacaaccact acacccagaa
     5581 gtccctgagc ctgagccccg gcggcagcgg tcttaatgat attttcgagg ctcagaagat
     5641 tgagtggcat gaacaccacc accatcacca ctaaggccgg cctgtagtca ctggcacaac
     5701 gcgttacccg gtaagccaat cgggtataca cggtcgtcat actgcagaca gggttcttct
     5761 actttgcaag atagtcttga gtagtaaaat aaatagatag agaaaatcag cctcgactgt
     5821 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5881 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcac aacactcaac
     5941 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     6001 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     6061 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6121 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6181 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     6241 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6301 cagaggccga ggccgcctct gcctctgagc tattccagaa
//
