LOCUS       sequence-368858-        6505 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 6505)
  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 6505)
  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-TNFSF4-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     243..263
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    381..492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     481..498
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(638..655)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(656..700)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1916..1935)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1948..1966)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1949..1966)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2176..2200)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2197..2400)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2226..2246)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2401..2780
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2947..2963
                     /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(3419..3436)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3419..3435)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3534..3551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3705..4293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3785..3804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4379..4398)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4404..4423)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4465..4556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4557..4625,4626..5417)
                     /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(4775..4794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5470..5591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5481..5500)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5535..5554
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5721..6320)
                     /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(5804..5824)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6301..6320
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6339..6386)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6434..6505,1..258))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6448..6505,1..78)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(6502..6505,1..16))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
       61 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
      121 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
      181 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      241 acaccctaac tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc
      301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
      361 gagatagggt tgagtgttgt gatgcaattt cctcatttta ttaggaaagg acagtgggag
      421 tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac
      481 tagaaggcac agtcgaggct gattttctct atctatttat tttactactc aagactatct
      541 tgcaaagtag aagaaccctg tctgcagtat gacgaccgtg tatacccgat tggcttaccg
      601 ggtaacgcgt tgtgccagtg actacaggcc ggccttagtg gtgatggtgg tggtgttcat
      661 gccactcaat cttctgagcc tcgaaaatat cattaagacc gctgccgccg gggctcaggc
      721 tcagggactt ctgggtgtag tggttgtgca gggcctcgtg catcacgctg cagctgaaca
      781 cgttgccctg ctgccacctg gacttgtcca cggtcagctt gctgtacagg aagaagctgc
      841 cgtcgctgtc cagcactggg ggggtggtct tgtagttgtt ctcgggctgg ccgttgctct
      901 cccactccac ggcgatatcg ctggggtaga agcccttcac cagacaggtc agggacacct
      961 ggttcttggt catctcctcc cggctggggg gcagggtgta cacctggggc tcccgtggct
     1021 ggcccttggc cttgctgatt gtcttttcga ttggggcagc cagggccttg ttggagactt
     1081 tgcacttgta ttctttgccg ttcagccagt cctggtgcag cacggtcagc acggacacca
     1141 ccctgtaggt gctgttgtac tgctcctctc tgggcttggt cttggcgttg tgcacctcca
     1201 cgccgtccac gtaccagttg aacttcacct ctgggtcctc gtgggacacg gccaccacca
     1261 cgcaggtcac ctcgggggtc ctgctgatca tcagggtgtc cttgggcttg ggggggaaca
     1321 ggaacacgga aggccctccc agcagttctg gagctgggca gggggggcag gtgtgggtct
     1381 tgtcgccgcc cggcgcgcca agcacacaga actcgccagg attctgatgg atcaggatca
     1441 gctcgccgcc gttcacatgg aaatcgtcca ggctggtgtt gtcggtggtc acgttcaggt
     1501 acaccttgtc cttgtaggtc aggctggcca ccatcaggct gttcacgctc cgcactttct
     1561 tcagctggaa caggggttcc tcgtccttct ggtagtgcag gctgatgttc acttcttggc
     1621 tgaagtagcc cttcaggctg atcaggtaga agccgtcgca gttgatgatc acgctgttgt
     1681 tctgcacctt catgatctcg tcctctttct ggctggtcag gatgaagccc ttctctttct
     1741 tgtactcggt gaactgcacc ttgatgctct ggattctggg gtatctgtga gacacttggg
     1801 cggccgcagc gaacaccagg cagaagatgt agctcagggc aatgatggtc ttcatggtgg
     1861 cgctagcagg cgcgttttcg atagaaccga ggtgcagttg gacctgggag tggacacctg
     1921 tggagagaaa ggcaaagtgg atgttattct atagtgtcac ctaaatcgta tgtgtatgat
     1981 acataaggtt atgtattaat tgtagccgcg ttctaacgaa gccaaggggg tgggcctata
     2041 gactctatag gcggtactta cgtcactctt ggcacgggga atccgcgttc caatgcaccg
     2101 ttcccggccg cggaggctgg atcggtcccg gtgtcttcta tggaggtcaa aacagcgtgg
     2161 atggcgtctc caggcgatct gacggttcac taaacgagct ctgcttatat agacctccca
     2221 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     2281 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     2341 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     2401 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     2461 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2521 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2581 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     2641 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     2701 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     2761 actagtcaat aatcaatgcc aacatggcgg tcatattgga catgagccaa tataaatgta
     2821 catattatga tatagataca acgtatgcaa tggccaatag ccaatattga tttatgctat
     2881 ataaccaatg aataatatgg ctaatggcca atattgagtc aacgccactg tgcatgaagc
     2941 tcgtaattgt tatccgctca caattaaggg cgacacaaaa tttattctaa atgataataa
     3001 atactgataa catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt
     3061 tgatatcaaa aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt
     3121 aacaaactag aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta
     3181 taggtgttca tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca
     3241 tttataaggt taaataattc tcatatatca agcaaagtga caggcgccct taaatattct
     3301 gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg
     3361 ttatttgcgg attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac
     3421 tggccgtcgt tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     3481 ggccttctgc ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc
     3541 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3601 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3661 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3721 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3781 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3841 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3901 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3961 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4021 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4081 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact
     4141 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4201 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4261 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4321 tctgacgctc agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagca
     4381 aggggtgtta tgagccatat tcaggtataa atgggctcgc gataatgttc agaattggtt
     4441 aattggttgt aacactgacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4501 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatatga
     4561 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4621 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4681 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4741 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4801 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4861 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4921 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4981 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5041 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5101 tagcgatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5161 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5221 cccttccggc tggctggttt attgctgata aatccggagc cggtgagcgt ggttctcgcg
     5281 gtatcatcgc agcgctgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5341 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5401 tgattaagca ttggtaatga ccatgattac gccaagctct agctagaggt cgacggtata
     5461 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     5521 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     5581 ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     5641 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     5701 tcgaaatctc gtagcacgtg tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
     5761 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     5821 ttgcggggcg cggatgtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     5881 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg
     5941 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     6001 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     6061 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     6121 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     6181 gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg
     6241 aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc
     6301 gtgggcttgt actcggtcat ggtttagttc ctcaccttgt cgtattatac tatgccgata
     6361 tactatgccg atgattaatt gtcaacacgt gctgatcaga tccgaaaatg gatatacaag
     6421 ctcccgggag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     6481 atagctcaga ggcagaggcg gcctc
//
