LOCUS       sequence-368749-        6643 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 6643)
  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 6643)
  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-FASLG-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     441..461
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    674..795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(711..730)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     765..784
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(848..1639,1640..1708))
                     /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     1471..1490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1709..1800)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1842..1861
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1867..1886
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1972..2560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2461..2480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2714..2731
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2829..2846
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2830..2846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3302..3318
                     /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        3485..3864
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3865..4068
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4019..4039
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4065..4089
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4299..4317
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4299..4316
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4330..4349
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5703..5747
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5748..5765
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5905..5922)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5911..6022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6140..6160)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6145..6474
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6325..6460
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6387..6406
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6522..6569
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(6588..6643,1..544)
                     /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(6588..6607)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
       61 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
      121 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
      181 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
      241 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
      301 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
      361 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
      421 cctggagaca tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
      481 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
      541 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
      601 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
      661 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
      721 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
      781 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
      841 atggtcatta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
      901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
      961 ctggccccag cgctgcgatg ataccgcgag aaccacgctc accggctccg gatttatcag
     1021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1141 tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     1681 gggcgacacg gaaatgttga atactcatat tcttcctttt tcaatattat tgaagcattt
     1741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     1801 taggggtcag tgttacaacc aattaaccaa ttctgaacat tatcgcgagc ccatttatac
     1861 ctgaatatgg ctcataacac cccttgctca tgaccaaaat cccttaacgt gagttacgcg
     1921 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1981 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2041 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2101 gcagatacca aatactgttc ttctagtgta gccgtagtta gcccaccact tcaagaactc
     2161 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2221 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2281 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2341 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2401 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2461 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2521 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2581 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2641 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2701 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa ggcgagagta gggaactgcc
     2761 aggcatcaaa ctaagcagaa ggcccctgac ggatggcctt tttgcgtttc tacaaactct
     2821 ttctgtgttg taaaacgacg gccagtctta agctcgggcc ccctgggcgg ttctgataac
     2881 gagtaatcgt taatccgcaa ataacgtaaa aacccgcttc ggcgggtttt tttatggggg
     2941 gagtttaggg aaagagcatt tgtcagaata tttaagggcg cctgtcactt tgcttgatat
     3001 atgagaatta tttaacctta taaatgagaa aaaagcaacg cactttaaat aagatacgtt
     3061 gctttttcga ttgatgaaca cctataatta aactattcat ctattattta tgattttttg
     3121 tatatacaat atttctagtt tgttaaagag aattaagaaa ataaatctcg aaaataataa
     3181 agggaaaatc agtttttgat atcaaaatta tacatgtcaa cgataataca aaatataata
     3241 caaactataa gatgttatca gtatttatta tcatttagaa taaattttgt gtcgccctta
     3301 attgtgagcg gataacaatt acgagcttca tgcacagtgg cgttgactca atattggcca
     3361 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     3421 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     3481 tgttggcatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     3541 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     3601 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     3661 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3721 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3781 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3841 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     3901 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     3961 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4021 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     4081 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     4141 cgatccagcc tccgcggccg ggaacggtgc attggaacgc ggattccccg tgccaagagt
     4201 gacgtaagta ccgcctatag agtctatagg cccaccccct tggcttcgtt agaacgcggc
     4261 tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaata
     4321 acatccactt tgcctttctc tccacaggtg tccactccca ggtccaactg cacctcggtt
     4381 ctatcgaaaa cgcgcctgct agcgccacca tgaagaccat cattgccctg agctacatct
     4441 tctgcctggt gttcgctgcg gccgcccaac tgttccatct gcaaaaagag ctggccgagc
     4501 tgcgggaaag cacctctcag atgcatacag ccagcagcct ggaaaagcag atcggacacc
     4561 ctagtcctcc acctgagaag aaagaactgc ggaaggtggc ccacctgacc ggcaagagca
     4621 atagcagatc catgcctctg gaatgggaag atacctacgg catcgtgctg ctgagcggcg
     4681 tgaagtataa gaaaggcggc ctggtcatca acgaaaccgg cctgtacttc gtgtacagca
     4741 aggtgtactt ccggggccag agctgcaaca atctgcctct gagccacaaa gtgtacatgc
     4801 ggaacagcaa gtaccctcag gacctggtca tgatggaagg caagatgatg agctactgca
     4861 ccaccggcca gatgtgggcc agatcttctt atctgggcgc cgtgttcaac ctgaccagcg
     4921 ccgatcacct gtacgtgaac gtgtccgagc tgagcctggt caacttcgag gaaagccaga
     4981 ccttctttgg cctgtacaag cttggcgcgc cgggcggcga caagacccac acctgccccc
     5041 cctgcccagc tccagaactg ctgggagggc cttccgtgtt cctgttcccc cccaagccca
     5101 aggacaccct gatgatcagc aggacccccg aggtgacctg cgtggtggtg gccgtgtccc
     5161 acgaggaccc agaggtgaag ttcaactggt acgtggacgg cgtggaggtg cacaacgcca
     5221 agaccaagcc cagagaggag cagtacaaca gcacctacag ggtggtgtcc gtgctgaccg
     5281 tgctgcacca ggactggctg aacggcaaag aatacaagtg caaagtctcc aacaaggccc
     5341 tggctgcccc aatcgaaaag acaatcagca aggccaaggg ccagccacgg gagccccagg
     5401 tgtacaccct gccccccagc cgggaggaga tgaccaagaa ccaggtgtcc ctgacctgtc
     5461 tggtgaaggg cttctacccc agcgatatcg ccgtggagtg ggagagcaac ggccagcccg
     5521 agaacaacta caagaccacc cccccagtgc tggacagcga cggcagcttc ttcctgtaca
     5581 gcaagctgac cgtggacaag tccaggtggc agcagggcaa cgtgttcagc tgcagcgtga
     5641 tgcacgaggc cctgcacaac cactacaccc agaagtccct gagcctgagc cccggcggca
     5701 gcggtcttaa tgatattttc gaggctcaga agattgagtg gcatgaacac caccaccatc
     5761 accactaagg ccggcctgta gtcactggca caacgcgtta cccggtaagc caatcgggta
     5821 tacacggtcg tcatactgca gacagggttc ttctactttg caagatagtc ttgagtagta
     5881 aaataaatag atagagaaaa tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     5941 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6001 aataaaatga ggaaattgca tcacaacact caaccctatc tcggtctatt cttttgattt
     6061 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6121 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     6181 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     6241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6301 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6361 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     6421 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6481 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     6541 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg accgagtaca
     6601 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggc
//
