LOCUS       sequence-368848-        6490 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 6490)
  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 6490)
  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-TNFSF18-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..188
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    306..417
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     406..423
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(563..580)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(581..625)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1826..1845)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1858..1876)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1859..1876)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2086..2110)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2107..2310)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2136..2156)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2311..2690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2857..2873
                     /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(3329..3346)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3329..3345)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3444..3461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3615..4203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3695..3714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4289..4308)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4314..4333)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4375..4466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4467..4535,4536..5327)
                     /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(4685..4704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5380..5501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5391..5410)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5445..5464
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5631..6230)
                     /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(5714..5734)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6211..6230
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6249..6296)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6344..6490,1..183))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6358..6490,1..3)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6412..6431)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
       61 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      121 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      181 cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
      241 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
      301 gttgtgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
      361 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
      421 aggctgattt tctctatcta tttattttac tactcaagac tatcttgcaa agtagaagaa
      481 ccctgtctgc agtatgacga ccgtgtatac ccgattggct taccgggtaa cgcgttgtgc
      541 cagtgactac aggccggcct tagtggtgat ggtggtggtg ttcatgccac tcaatcttct
      601 gagcctcgaa aatatcatta agaccgctgc cgccggggct caggctcagg gacttctggg
      661 tgtagtggtt gtgcagggcc tcgtgcatca cgctgcagct gaacacgttg ccctgctgcc
      721 acctggactt gtccacggtc agcttgctgt acaggaagaa gctgccgtcg ctgtccagca
      781 ctgggggggt ggtcttgtag ttgttctcgg gctggccgtt gctctcccac tccacggcga
      841 tatcgctggg gtagaagccc ttcaccagac aggtcaggga cacctggttc ttggtcatct
      901 cctcccggct ggggggcagg gtgtacacct ggggctcccg tggctggccc ttggccttgc
      961 tgattgtctt ttcgattggg gcagccaggg ccttgttgga gactttgcac ttgtattctt
     1021 tgccgttcag ccagtcctgg tgcagcacgg tcagcacgga caccaccctg taggtgctgt
     1081 tgtactgctc ctctctgggc ttggtcttgg cgttgtgcac ctccacgccg tccacgtacc
     1141 agttgaactt cacctctggg tcctcgtggg acacggccac caccacgcag gtcacctcgg
     1201 gggtcctgct gatcatcagg gtgtccttgg gcttgggggg gaacaggaac acggaaggcc
     1261 ctcccagcag ttctggagct gggcaggggg ggcaggtgtg ggtcttgtcg ccgcccggcg
     1321 cgccagagat gaactgagga ttggccagca ggatgatgcc ccagtaggtg ttgttcttca
     1381 gcacctggtg ctcgctgttg aagatcaggt cgatggtatc gcccacgtgc agctcatagg
     1441 tgccgccgac gttctggatc ttgctcttgt tggtcagtgt ctggatcatg tccttgttct
     1501 tgtacagccg cacttcgaaa ggggccacat cgttgtagtt ggcgttaggg gccacctggc
     1561 catagatcag gtacaggccg ttctgcagga tttccagctt ccagtcggac accttgttca
     1621 cgcaaggagg ctcgctagag gccatctgcc acttagaagg cagagggccg aacttggcca
     1681 tgcaaggttc tttggcggtt tccagttggg cggccgcagc gaacaccagg cagaagatgt
     1741 agctcagggc aatgatggtc ttcatggtgg cgctagcagg cgcgttttcg atagaaccga
     1801 ggtgcagttg gacctgggag tggacacctg tggagagaaa ggcaaagtgg atgttattct
     1861 atagtgtcac ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg
     1921 ttctaacgaa gccaaggggg tgggcctata gactctatag gcggtactta cgtcactctt
     1981 ggcacgggga atccgcgttc caatgcaccg ttcccggccg cggaggctgg atcggtcccg
     2041 gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac
     2101 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     2161 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     2221 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     2281 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     2341 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     2401 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     2461 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     2521 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     2581 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     2641 taatgacccc gtaattgatt actattaata actagtcaat aatcaatgcc aacatggcgg
     2701 tcatattgga catgagccaa tataaatgta catattatga tatagataca acgtatgcaa
     2761 tggccaatag ccaatattga tttatgctat ataaccaatg aataatatgg ctaatggcca
     2821 atattgagtc aacgccactg tgcatgaagc tcgtaattgt tatccgctca caattaaggg
     2881 cgacacaaaa tttattctaa atgataataa atactgataa catcttatag tttgtattat
     2941 attttgtatt atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta
     3001 ttttcgagat ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa
     3061 atcataaata atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta
     3121 tcttatttaa agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca
     3181 agcaaagtga caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca
     3241 taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc
     3301 agaaccgccc agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt
     3361 tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt
     3421 tccctactct cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3481 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3541 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4021 ttgaagtggt gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta
     4261 actcacgtta agggattttg gtcatgagca aggggtgtta tgagccatat tcaggtataa
     4321 atgggctcgc gataatgttc agaattggtt aattggttgt aacactgacc cctatttgtt
     4381 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4441 ttcaataata ttgaaaaagg aagaatatga gtattcaaca tttccgtgtc gcccttattc
     4501 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4561 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4621 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4681 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4741 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4801 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4861 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4921 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4981 caaacgacga gcgtgacacc acgatgcctg tagcgatggc aacaacgttg cgcaaactat
     5041 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5101 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5161 aatccggagc cggtgagcgt ggttctcgcg gtatcatcgc agcgctgggg ccagatggta
     5221 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5281 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaatga ccatgattac
     5341 gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg
     5401 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     5461 ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc
     5521 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     5581 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtagcacgtg tcaggcaccg
     5641 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg
     5701 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggatgtctc caggaaggcg
     5761 ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc
     5821 ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc
     5881 cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac
     5941 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg
     6001 gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg
     6061 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg
     6121 acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg
     6181 gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtttagttc
     6241 ctcaccttgt cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt
     6301 gctgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa aagcctaggc
     6361 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg gcctcggcct
     6421 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     6481 ttaggggcgg
//
