LOCUS       sequence-420969-        6679 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 6679)
  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 6679)
  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-CD48-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     87..107
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    225..336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     325..342
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(482..499)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(500..544)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1934..1953)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1966..1984)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1967..1984)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2194..2218)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2215..2418)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2244..2264)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2419..2798
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2965..2981
                     /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(3437..3454)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3437..3453)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3552..3569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3723..4311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3803..3822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4397..4416)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4422..4441)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4483..4574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4575..4643,4644..5435)
                     /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(4793..4812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5488..5609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5499..5518)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5553..5572
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5739..6338)
                     /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(5822..5842)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6319..6338
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6357..6404)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6452..6679,1..102))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6466..6601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6520..6539)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
       61 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gaattaattc
      121 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
      181 ccttataaat caaaagaata gaccgagata gggttgagtg ttgtgatgca atttcctcat
      241 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
      301 ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatttt ctctatctat
      361 ttattttact actcaagact atcttgcaaa gtagaagaac cctgtctgca gtatgacgac
      421 cgtgtatacc cgattggctt accgggtaac gcgttgtgcc agtgactaca ggccggcctt
      481 agtggtgatg gtggtggtgt tcatgccact caatcttctg agcctcgaaa atatcattaa
      541 gaccgctgcc gccggggctc aggctcaggg acttctgggt gtagtggttg tgcagggcct
      601 cgtgcatcac gctgcagctg aacacgttgc cctgctgcca cctggacttg tccacggtca
      661 gcttgctgta caggaagaag ctgccgtcgc tgtccagcac tgggggggtg gtcttgtagt
      721 tgttctcggg ctggccgttg ctctcccact ccacggcgat atcgctgggg tagaagccct
      781 tcaccagaca ggtcagggac acctggttct tggtcatctc ctcccggctg gggggcaggg
      841 tgtacacctg gggctcccgt ggctggccct tggccttgct gattgtcttt tcgattgggg
      901 cagccagggc cttgttggag actttgcact tgtattcttt gccgttcagc cagtcctggt
      961 gcagcacggt cagcacggac accaccctgt aggtgctgtt gtactgctcc tctctgggct
     1021 tggtcttggc gttgtgcacc tccacgccgt ccacgtacca gttgaacttc acctctgggt
     1081 cctcgtggga cacggccacc accacgcagg tcacctcggg ggtcctgctg atcatcaggg
     1141 tgtccttggg cttggggggg aacaggaaca cggaaggccc tcccagcagt tctggagctg
     1201 ggcagggggg gcaggtgtgg gtcttgtcgc cgcccggcgc gccaagtgtg catggagggc
     1261 tcagacacac ggtgccattc ttgctggaca cgctgttgga cacctggcat gtgtagcacc
     1321 ggctgtagtt gtggggcatc agtgtggttt ccagcacgct gttctgcagt tcttttggaa
     1381 agggccgctt gtcgccgtac caagtgtagt tcacgctctc gccagggatc acgcagctca
     1441 gcttcaggta gcagttgtcg tccatgtcct caatcttctc gatcttgatc acaggcttag
     1501 gcacagggtc cagcacctgg agcttgatct tccactcttg ctcgttgccg gttttcttca
     1561 gcacccgcat gatgtaggtg ctgttgtcct ctttctgcac cttgctgata tacagggcgc
     1621 cagactgagg atcgagccgc actctgccct tgaacttgct ctcgaagtac ttggacttcc
     1681 ggctgtccca ttccacgatt ttctggtcga aggtgtagaa ccaggtcagc tgcttgtagt
     1741 tctcaggcag gctctcagag atgttcaggg tcacgttgct gccggacacc actgtcatat
     1801 gcaccagatg tccttgggcg gccgcagcga acaccaggca gaagatgtag ctcagggcaa
     1861 tgatggtctt catggtggcg ctagcaggcg cgttttcgat agaaccgagg tgcagttgga
     1921 cctgggagtg gacacctgtg gagagaaagg caaagtggat gttattctat agtgtcacct
     1981 aaatcgtatg tgtatgatac ataaggttat gtattaattg tagccgcgtt ctaacgaagc
     2041 caagggggtg ggcctataga ctctataggc ggtacttacg tcactcttgg cacggggaat
     2101 ccgcgttcca atgcaccgtt cccggccgcg gaggctggat cggtcccggt gtcttctatg
     2161 gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct
     2221 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     2281 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     2341 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     2401 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     2461 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     2521 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     2581 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     2641 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     2701 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     2761 aattgattac tattaataac tagtcaataa tcaatgccaa catggcggtc atattggaca
     2821 tgagccaata taaatgtaca tattatgata tagatacaac gtatgcaatg gccaatagcc
     2881 aatattgatt tatgctatat aaccaatgaa taatatggct aatggccaat attgagtcaa
     2941 cgccactgtg catgaagctc gtaattgtta tccgctcaca attaagggcg acacaaaatt
     3001 tattctaaat gataataaat actgataaca tcttatagtt tgtattatat tttgtattat
     3061 cgttgacatg tataattttg atatcaaaaa ctgattttcc ctttattatt ttcgagattt
     3121 attttcttaa ttctctttaa caaactagaa atattgtata tacaaaaaat cataaataat
     3181 agatgaatag tttaattata ggtgttcatc aatcgaaaaa gcaacgtatc ttatttaaag
     3241 tgcgttgctt ttttctcatt tataaggtta aataattctc atatatcaag caaagtgaca
     3301 ggcgccctta aatattctga caaatgctct ttccctaaac tccccccata aaaaaacccg
     3361 ccgaagcggg tttttacgtt atttgcggat taacgattac tcgttatcag aaccgcccag
     3421 ggggcccgag cttaagactg gccgtcgttt tacaacacag aaagagtttg tagaaacgca
     3481 aaaaggccat ccgtcagggg ccttctgctt agtttgatgc ctggcagttc cctactctcg
     3541 ccttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3901 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4141 gctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4261 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4321 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag
     4381 ggattttggt catgagcaag gggtgttatg agccatattc aggtataaat gggctcgcga
     4441 taatgttcag aattggttaa ttggttgtaa cactgacccc tatttgttta tttttctaaa
     4501 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4561 gaaaaaggaa gaatatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4621 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4681 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4741 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4801 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4861 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4921 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4981 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5041 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5101 gtgacaccac gatgcctgta gcgatggcaa caacgttgcg caaactatta actggcgaac
     5161 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5221 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tccggagccg
     5281 gtgagcgtgg ttctcgcggt atcatcgcag cgctggggcc agatggtaag ccctcccgta
     5341 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5401 ctgagatagg tgcctcactg attaagcatt ggtaatgacc atgattacgc caagctctag
     5461 ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa
     5521 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5581 taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc
     5641 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     5701 tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc aggcaccggg cttgcgggtc
     5761 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     5821 cgctcgtaga aggggaggtt gcggggcgcg gatgtctcca ggaaggcggg caccccggcg
     5881 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     5941 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     6001 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     6061 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     6121 accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     6181 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc
     6241 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg
     6301 cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg tttagttcct caccttgtcg
     6361 tattatacta tgccgatata ctatgccgat gattaattgt caacacgtgc tgatcagatc
     6421 cgaaaatgga tatacaagct cccgggagct ttttgcaaaa gcctaggcct ccaaaaaagc
     6481 ctcctcacta cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa
     6541 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     6601 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     6661 ttctgcctgc tggggagcc
//
