LOCUS       sequence-368735-        6610 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 6610)
  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 6610)
  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-CLEC4A-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     160..180
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    298..409
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     398..415
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(555..572)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(573..617)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1938..1957)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1970..1988)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1971..1988)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2198..2222)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2219..2422)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2248..2268)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2423..2802
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2969..2985
                     /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(3441..3458)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3441..3457)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3556..3573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3727..4315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3807..3826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4401..4420)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4426..4445)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4487..4578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4579..4647,4648..5439)
                     /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(4797..4816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5492..5613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5557..5576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5743..6342)
                     /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(5826..5846)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6323..6342
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6361..6408)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6456..6610,1..175))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6470..6605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6524..6543)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
       61 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
      121 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
      181 acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
      241 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgtgat
      301 gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa
      361 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt cgaggctgat
      421 tttctctatc tatttatttt actactcaag actatcttgc aaagtagaag aaccctgtct
      481 gcagtatgac gaccgtgtat acccgattgg cttaccgggt aacgcgttgt gccagtgact
      541 acaggccggc cttagtggtg atggtggtgg tgttcatgcc actcaatctt ctgagcctcg
      601 aaaatatcat taagaccgct gccgccgggg ctcaggctca gggacttctg ggtgtagtgg
      661 ttgtgcaggg cctcgtgcat cacgctgcag ctgaacacgt tgccctgctg ccacctggac
      721 ttgtccacgg tcagcttgct gtacaggaag aagctgccgt cgctgtccag cactgggggg
      781 gtggtcttgt agttgttctc gggctggccg ttgctctccc actccacggc gatatcgctg
      841 gggtagaagc ccttcaccag acaggtcagg gacacctggt tcttggtcat ctcctcccgg
      901 ctggggggca gggtgtacac ctggggctcc cgtggctggc ccttggcctt gctgattgtc
      961 ttttcgattg gggcagccag ggccttgttg gagactttgc acttgtattc tttgccgttc
     1021 agccagtcct ggtgcagcac ggtcagcacg gacaccaccc tgtaggtgct gttgtactgc
     1081 tcctctctgg gcttggtctt ggcgttgtgc acctccacgc cgtccacgta ccagttgaac
     1141 ttcacctctg ggtcctcgtg ggacacggcc accaccacgc aggtcacctc gggggtcctg
     1201 ctgatcatca gggtgtcctt gggcttgggg gggaacagga acacggaagg ccctcccagc
     1261 agttctggag ctgggcaggg ggggcaggtg tgggtcttgt cgccgcccgg cgcgccaagg
     1321 tggatcttca tcatctcgca cacgcttctc tgagggccca gacagttcac gtcgttccag
     1381 ccccatctct tagggctctt gcggaagttc agcaccacgc atctctcgtt aggatcgctt
     1441 ggctctctgg ggtgccagaa ggtgctgctc tcgttgtaag gtgtctggtc gacccactgc
     1501 cagtgtctct gtccttcagg atcagacagg cccacgaagt aggcggattc ctcttgcagg
     1561 ttttggaaga tgaagtcctg ttcctcttgg gtgttgatga ccagcagatg ggcttccatc
     1621 cgggcacagt ccttctcaga gtcctgccaa gaggcgctct cggtgctgat gaagtagcag
     1681 ttgctgctga aggacttcca gttcttggga cagcaggacc aggctgtctc ttccacgggc
     1741 atgtttttct tcacgcactc gagggttgtg tgcaccagtt ctttggtggt tttcttttcc
     1801 agcagctggg agtatttctg ggcggccgca gcgaacacca ggcagaagat gtagctcagg
     1861 gcaatgatgg tcttcatggt ggcgctagca ggcgcgtttt cgatagaacc gaggtgcagt
     1921 tggacctggg agtggacacc tgtggagaga aaggcaaagt ggatgttatt ctatagtgtc
     1981 acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg
     2041 aagccaaggg ggtgggccta tagactctat aggcggtact tacgtcactc ttggcacggg
     2101 gaatccgcgt tccaatgcac cgttcccggc cgcggaggct ggatcggtcc cggtgtcttc
     2161 tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc actaaacgag
     2221 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     2281 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     2341 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     2401 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     2461 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     2521 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     2581 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     2641 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     2701 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     2761 ccgtaattga ttactattaa taactagtca ataatcaatg ccaacatggc ggtcatattg
     2821 gacatgagcc aatataaatg tacatattat gatatagata caacgtatgc aatggccaat
     2881 agccaatatt gatttatgct atataaccaa tgaataatat ggctaatggc caatattgag
     2941 tcaacgccac tgtgcatgaa gctcgtaatt gttatccgct cacaattaag ggcgacacaa
     3001 aatttattct aaatgataat aaatactgat aacatcttat agtttgtatt atattttgta
     3061 ttatcgttga catgtataat tttgatatca aaaactgatt ttccctttat tattttcgag
     3121 atttattttc ttaattctct ttaacaaact agaaatattg tatatacaaa aaatcataaa
     3181 taatagatga atagtttaat tataggtgtt catcaatcga aaaagcaacg tatcttattt
     3241 aaagtgcgtt gcttttttct catttataag gttaaataat tctcatatat caagcaaagt
     3301 gacaggcgcc cttaaatatt ctgacaaatg ctctttccct aaactccccc cataaaaaaa
     3361 cccgccgaag cgggttttta cgttatttgc ggattaacga ttactcgtta tcagaaccgc
     3421 ccagggggcc cgagcttaag actggccgtc gttttacaac acagaaagag tttgtagaaa
     3481 cgcaaaaagg ccatccgtca ggggccttct gcttagtttg atgcctggca gttccctact
     3541 ctcgccttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4141 gtgggctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt
     4381 taagggattt tggtcatgag caaggggtgt tatgagccat attcaggtat aaatgggctc
     4441 gcgataatgt tcagaattgg ttaattggtt gtaacactga cccctatttg tttatttttc
     4501 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4561 tattgaaaaa ggaagaatat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4621 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4681 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4741 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4801 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4861 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4921 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4981 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5041 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5101 gagcgtgaca ccacgatgcc tgtagcgatg gcaacaacgt tgcgcaaact attaactggc
     5161 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5221 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatccgga
     5281 gccggtgagc gtggttctcg cggtatcatc gcagcgctgg ggccagatgg taagccctcc
     5341 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5401 atcgctgaga taggtgcctc actgattaag cattggtaat gaccatgatt acgccaagct
     5461 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc
     5521 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5581 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
     5641 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     5701 ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg tgtcaggcac cgggcttgcg
     5761 ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc
     5821 gagccgctcg tagaagggga ggttgcgggg cgcggatgtc tccaggaagg cgggcacccc
     5881 ggcgcgctcg gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg
     5941 gtggtcgggc gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg
     6001 cgccaggagg ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat
     6061 gcgcgggccg atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac
     6121 cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt
     6181 gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg
     6241 gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc
     6301 gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc atggtttagt tcctcacctt
     6361 gtcgtattat actatgccga tatactatgc cgatgattaa ttgtcaacac gtgctgatca
     6421 gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa
     6481 aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa
     6541 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     6601 gggatgggcg
//
