LOCUS       sequence-368599-        6508 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 6508)
  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 6508)
  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-CD28-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    351..462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     451..468
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(608..625)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(626..670)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1889..1908)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1921..1939)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1922..1939)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2149..2173)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2170..2373)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2199..2219)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2374..2753
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2920..2936
                     /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(3392..3409)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3392..3408)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3507..3524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3678..4266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3758..3777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4352..4371)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4377..4396)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4438..4529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4530..4598,4599..5390)
                     /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(4748..4767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5443..5564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5454..5473)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5508..5527
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5694..6293)
                     /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(5777..5797)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6274..6293
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6312..6359)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6407..6508,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6421..6508,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6475..6494)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
      241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt gatgcaattt
      361 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
      421 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gattttctct
      481 atctatttat tttactactc aagactatct tgcaaagtag aagaaccctg tctgcagtat
      541 gacgaccgtg tatacccgat tggcttaccg ggtaacgcgt tgtgccagtg actacaggcc
      601 ggccttagtg gtgatggtgg tggtgttcat gccactcaat cttctgagcc tcgaaaatat
      661 cattaagacc gctgccgccg gggctcaggc tcagggactt ctgggtgtag tggttgtgca
      721 gggcctcgtg catcacgctg cagctgaaca cgttgccctg ctgccacctg gacttgtcca
      781 cggtcagctt gctgtacagg aagaagctgc cgtcgctgtc cagcactggg ggggtggtct
      841 tgtagttgtt ctcgggctgg ccgttgctct cccactccac ggcgatatcg ctggggtaga
      901 agcccttcac cagacaggtc agggacacct ggttcttggt catctcctcc cggctggggg
      961 gcagggtgta cacctggggc tcccgtggct ggcccttggc cttgctgatt gtcttttcga
     1021 ttggggcagc cagggccttg ttggagactt tgcacttgta ttctttgccg ttcagccagt
     1081 cctggtgcag cacggtcagc acggacacca ccctgtaggt gctgttgtac tgctcctctc
     1141 tgggcttggt cttggcgttg tgcacctcca cgccgtccac gtaccagttg aacttcacct
     1201 ctgggtcctc gtgggacacg gccaccacca cgcaggtcac ctcgggggtc ctgctgatca
     1261 tcagggtgtc cttgggcttg ggggggaaca ggaacacgga aggccctccc agcagttctg
     1321 gagctgggca gggggggcag gtgtgggtct tgtcgccgcc cggcgcgcca ggcttgctag
     1381 gtccagggaa cagtggagaa ggacacaggt gcttgccctt cacgtggatg atggtgccgt
     1441 tggacttctc gttgtccagg taaggaggag ggtacatcac ttcgatcttg cagaagtaga
     1501 tatcggtctg gttcacgtac aggttttgca ggtagaaggt cacgctctcg ttgcccagct
     1561 tgccgtcaca attgaagccg gtcttgctgt agacctgcag ctgctggctg taattgccgt
     1621 acaccacgca cacttccacg gcagaatcca gtcctttgtg caggctggcc cggaactctc
     1681 tgctgaacag gttgtagctg tacttgcagc tcaggttcac ggcgttgtcg taggccacca
     1741 gcataggaga ctgcttgacc aggatcttgt tggcggccgc agcgaacacc aggcagaaga
     1801 tgtagctcag ggcaatgatg gtcttcatgg tggcgctagc aggcgcgttt tcgatagaac
     1861 cgaggtgcag ttggacctgg gagtggacac ctgtggagag aaaggcaaag tggatgttat
     1921 tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc
     1981 gcgttctaac gaagccaagg gggtgggcct atagactcta taggcggtac ttacgtcact
     2041 cttggcacgg ggaatccgcg ttccaatgca ccgttcccgg ccgcggaggc tggatcggtc
     2101 ccggtgtctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga tctgacggtt
     2161 cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     2221 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2281 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     2341 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     2401 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     2461 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     2521 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     2581 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     2641 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     2701 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gccaacatgg
     2761 cggtcatatt ggacatgagc caatataaat gtacatatta tgatatagat acaacgtatg
     2821 caatggccaa tagccaatat tgatttatgc tatataacca atgaataata tggctaatgg
     2881 ccaatattga gtcaacgcca ctgtgcatga agctcgtaat tgttatccgc tcacaattaa
     2941 gggcgacaca aaatttattc taaatgataa taaatactga taacatctta tagtttgtat
     3001 tatattttgt attatcgttg acatgtataa ttttgatatc aaaaactgat tttcccttta
     3061 ttattttcga gatttatttt cttaattctc tttaacaaac tagaaatatt gtatatacaa
     3121 aaaatcataa ataatagatg aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac
     3181 gtatcttatt taaagtgcgt tgcttttttc tcatttataa ggttaaataa ttctcatata
     3241 tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat gctctttccc taaactcccc
     3301 ccataaaaaa acccgccgaa gcgggttttt acgttatttg cggattaacg attactcgtt
     3361 atcagaaccg cccagggggc ccgagcttaa gactggccgt cgttttacaa cacagaaaga
     3421 gtttgtagaa acgcaaaaag gccatccgtc aggggccttc tgcttagttt gatgcctggc
     3481 agttccctac tctcgccttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3541 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4081 ttcttgaagt ggtgggctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc
     4321 gtaactcacg ttaagggatt ttggtcatga gcaaggggtg ttatgagcca tattcaggta
     4381 taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt
     4441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4501 tgcttcaata atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta
     4561 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4621 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4681 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4741 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4801 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4861 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4921 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4981 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5041 taccaaacga cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac
     5101 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5161 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5221 ataaatccgg agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg
     5281 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5341 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa tgaccatgat
     5401 tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt
     5461 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     5521 ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac
     5581 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     5641 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtagcac gtgtcaggca
     5701 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     5761 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggatgt ctccaggaag
     5821 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     5881 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     5941 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     6001 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     6061 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     6121 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga
     6181 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
     6241 ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catggtttag
     6301 ttcctcacct tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca
     6361 cgtgctgatc agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta
     6421 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg
     6481 cctctgcata aataaaaaaa attagtca
//
