LOCUS       sequence-368604-        6517 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 6517)
  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 6517)
  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-TNFRSF18-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     238..258
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    376..487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     476..493
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(633..650)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(651..695)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1923..1942)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1955..1973)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1956..1973)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2183..2207)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2204..2407)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2233..2253)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2408..2787
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2954..2970
                     /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(3426..3443)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3426..3442)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3541..3558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3712..4300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3792..3811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4386..4405)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4411..4430)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4472..4563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4564..4632,4633..5424)
                     /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(4782..4801)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5477..5598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5488..5507)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5542..5561
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5728..6327)
                     /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(5811..5831)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6308..6327
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6346..6393)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6441..6517,1..253))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6455..6517,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(6509..6517,1..11))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
       61 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
      121 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
      181 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
      241 ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt
      301 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      361 agggttgagt gttgtgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
      421 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
      481 ggcacagtcg aggctgattt tctctatcta tttattttac tactcaagac tatcttgcaa
      541 agtagaagaa ccctgtctgc agtatgacga ccgtgtatac ccgattggct taccgggtaa
      601 cgcgttgtgc cagtgactac aggccggcct tagtggtgat ggtggtggtg ttcatgccac
      661 tcaatcttct gagcctcgaa aatatcatta agaccgctgc cgccggggct caggctcagg
      721 gacttctggg tgtagtggtt gtgcagggcc tcgtgcatca cgctgcagct gaacacgttg
      781 ccctgctgcc acctggactt gtccacggtc agcttgctgt acaggaagaa gctgccgtcg
      841 ctgtccagca ctgggggggt ggtcttgtag ttgttctcgg gctggccgtt gctctcccac
      901 tccacggcga tatcgctggg gtagaagccc ttcaccagac aggtcaggga cacctggttc
      961 ttggtcatct cctcccggct ggggggcagg gtgtacacct ggggctcccg tggctggccc
     1021 ttggccttgc tgattgtctt ttcgattggg gcagccaggg ccttgttgga gactttgcac
     1081 ttgtattctt tgccgttcag ccagtcctgg tgcagcacgg tcagcacgga caccaccctg
     1141 taggtgctgt tgtactgctc ctctctgggc ttggtcttgg cgttgtgcac ctccacgccg
     1201 tccacgtacc agttgaactt cacctctggg tcctcgtggg acacggccac caccacgcag
     1261 gtcacctcgg gggtcctgct gatcatcagg gtgtccttgg gcttgggggg gaacaggaac
     1321 acggaaggcc ctcccagcag ttctggagct gggcaggggg ggcaggtgtg ggtcttgtcg
     1381 ccgcccggcg cgccaggttc ggcaggagga gagcctggaa cacacacggc attgtgggtc
     1441 ttgttgccgg gaaacacggt cagaaagccg aactgggtgc agtctgtcca aggcttgcag
     1501 tgtccctcgt gtccgccaga aaatgtgccg ctggcgcaat cgatgcactg gaagccgaag
     1561 ctgaacttgc cctgagactg cacgccctgt ccaggtggac aaggatggtg tctgcatgtg
     1621 gtgcagcaag gatcgccgca gtggaactca ggctgcacgc acatacagtc ccactcgcta
     1681 cagcattcct cgccagggta gtccctgcaa catctggtgg tgtgcactct gcagcatctg
     1741 gcatcggtgc ctgttccaag cagcagtctg cctggtccac atccaggtcc acctgtaggt
     1801 ctttgggcgg ccgcagcgaa caccaggcag aagatgtagc tcagggcaat gatggtcttc
     1861 atggtggcgc tagcaggcgc gttttcgata gaaccgaggt gcagttggac ctgggagtgg
     1921 acacctgtgg agagaaaggc aaagtggatg ttattctata gtgtcaccta aatcgtatgt
     1981 gtatgataca taaggttatg tattaattgt agccgcgttc taacgaagcc aagggggtgg
     2041 gcctatagac tctataggcg gtacttacgt cactcttggc acggggaatc cgcgttccaa
     2101 tgcaccgttc ccggccgcgg aggctggatc ggtcccggtg tcttctatgg aggtcaaaac
     2161 agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg cttatataga
     2221 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     2281 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     2341 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     2401 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     2461 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     2521 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     2581 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     2641 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     2701 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     2761 attaataact agtcaataat caatgccaac atggcggtca tattggacat gagccaatat
     2821 aaatgtacat attatgatat agatacaacg tatgcaatgg ccaatagcca atattgattt
     2881 atgctatata accaatgaat aatatggcta atggccaata ttgagtcaac gccactgtgc
     2941 atgaagctcg taattgttat ccgctcacaa ttaagggcga cacaaaattt attctaaatg
     3001 ataataaata ctgataacat cttatagttt gtattatatt ttgtattatc gttgacatgt
     3061 ataattttga tatcaaaaac tgattttccc tttattattt tcgagattta ttttcttaat
     3121 tctctttaac aaactagaaa tattgtatat acaaaaaatc ataaataata gatgaatagt
     3181 ttaattatag gtgttcatca atcgaaaaag caacgtatct tatttaaagt gcgttgcttt
     3241 tttctcattt ataaggttaa ataattctca tatatcaagc aaagtgacag gcgcccttaa
     3301 atattctgac aaatgctctt tccctaaact ccccccataa aaaaacccgc cgaagcgggt
     3361 ttttacgtta tttgcggatt aacgattact cgttatcaga accgcccagg gggcccgagc
     3421 ttaagactgg ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa aaaggccatc
     3481 cgtcaggggc cttctgctta gtttgatgcc tggcagttcc ctactctcgc cttccgcttc
     3541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3901 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggg ctaactacgg
     4141 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4321 tacggggtct gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg gattttggtc
     4381 atgagcaagg ggtgttatga gccatattca ggtataaatg ggctcgcgat aatgttcaga
     4441 attggttaat tggttgtaac actgacccct atttgtttat ttttctaaat acattcaaat
     4501 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4561 aatatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4621 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4681 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4741 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4801 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4861 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4921 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4981 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5041 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5101 atgcctgtag cgatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5161 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5221 cgctcggccc ttccggctgg ctggtttatt gctgataaat ccggagccgg tgagcgtggt
     5281 tctcgcggta tcatcgcagc gctggggcca gatggtaagc cctcccgtat cgtagttatc
     5341 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5401 gcctcactga ttaagcattg gtaatgacca tgattacgcc aagctctagc tagaggtcga
     5461 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     5521 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     5581 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     5641 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     5701 ggtggaatcg aaatctcgta gcacgtgtca ggcaccgggc ttgcgggtca tgcaccaggt
     5761 gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa
     5821 ggggaggttg cggggcgcgg atgtctccag gaaggcgggc accccggcgc gctcggccgc
     5881 ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac
     5941 gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc
     6001 catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc
     6061 ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc
     6121 gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg
     6181 cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta
     6241 gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag
     6301 gcgcaccgtg ggcttgtact cggtcatggt ttagttcctc accttgtcgt attatactat
     6361 gccgatatac tatgccgatg attaattgtc aacacgtgct gatcagatcc gaaaatggat
     6421 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     6481 ttctggaata gctcagaggc agaggcggcc tcggcct
//
