LOCUS       sequence-368506-        6268 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 6268)
  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 6268)
  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-TNFRSF17-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     140..159
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(178..225)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(273..602)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      287..422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(341..360)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     587..607
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    725..836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     825..842
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(982..999)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1000..1044)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2023..2042)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2055..2073)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2056..2073)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2283..2307)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2304..2507)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2333..2353)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2508..2887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3054..3070
                     /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(3526..3543)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3526..3542)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3641..3658)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3812..4400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3892..3911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4486..4505)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4511..4530)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4572..4663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4664..4732,4733..5524)
                     /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(4882..4901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5577..5698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5588..5607)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5642..5661
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5828..6268,1..159))
                     /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(5911..5931)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gaagagttct tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg
       61 cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc
      121 gcgggtggcg aggcgcaccg tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc
      181 gtattatact atgccgatat actatgccga tgattaattg tcaacacgtg ctgatcagat
      241 ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag
      301 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
      361 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
      421 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
      481 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
      541 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
      601 acattccaca gaattaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
      661 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      721 ttgtgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
      781 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
      841 ggctgatttt ctctatctat ttattttact actcaagact atcttgcaaa gtagaagaac
      901 cctgtctgca gtatgacgac cgtgtatacc cgattggctt accgggtaac gcgttgtgcc
      961 agtgactaca ggccggcctt agtggtgatg gtggtggtgt tcatgccact caatcttctg
     1021 agcctcgaaa atatcattaa gaccgctgcc gccggggctc aggctcaggg acttctgggt
     1081 gtagtggttg tgcagggcct cgtgcatcac gctgcagctg aacacgttgc cctgctgcca
     1141 cctggacttg tccacggtca gcttgctgta caggaagaag ctgccgtcgc tgtccagcac
     1201 tgggggggtg gtcttgtagt tgttctcggg ctggccgttg ctctcccact ccacggcgat
     1261 atcgctgggg tagaagccct tcaccagaca ggtcagggac acctggttct tggtcatctc
     1321 ctcccggctg gggggcaggg tgtacacctg gggctcccgt ggctggccct tggccttgct
     1381 gattgtcttt tcgattgggg cagccagggc cttgttggag actttgcact tgtattcttt
     1441 gccgttcagc cagtcctggt gcagcacggt cagcacggac accaccctgt aggtgctgtt
     1501 gtactgctcc tctctgggct tggtcttggc gttgtgcacc tccacgccgt ccacgtacca
     1561 gttgaacttc acctctgggt cctcgtggga cacggccacc accacgcagg tcacctcggg
     1621 ggtcctgctg atcatcaggg tgtccttggg cttggggggg aacaggaaca cggaaggccc
     1681 tcccagcagt tctggagctg ggcagggggg gcaggtgtgg gtcttgtcgc cgcccggcgc
     1741 gccagcattg gtgcccttca cgctattggt cacgctggcg ttgcagtatc tctggcaggt
     1801 cagtggaggg gtgttgctgc tacatctcag ctggcaggga atgcaggcgt gcagcaggct
     1861 atcgaagtac tcgttctggc tacactggcc agccatctga agcatggcgg ccgcagcgaa
     1921 caccaggcag aagatgtagc tcagggcaat gatggtcttc atggtggcgc tagcaggcgc
     1981 gttttcgata gaaccgaggt gcagttggac ctgggagtgg acacctgtgg agagaaaggc
     2041 aaagtggatg ttattctata gtgtcaccta aatcgtatgt gtatgataca taaggttatg
     2101 tattaattgt agccgcgttc taacgaagcc aagggggtgg gcctatagac tctataggcg
     2161 gtacttacgt cactcttggc acggggaatc cgcgttccaa tgcaccgttc ccggccgcgg
     2221 aggctggatc ggtcccggtg tcttctatgg aggtcaaaac agcgtggatg gcgtctccag
     2281 gcgatctgac ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta
     2341 ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt
     2401 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     2461 tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg
     2521 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     2581 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     2641 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     2701 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     2761 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     2821 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     2881 caatgccaac atggcggtca tattggacat gagccaatat aaatgtacat attatgatat
     2941 agatacaacg tatgcaatgg ccaatagcca atattgattt atgctatata accaatgaat
     3001 aatatggcta atggccaata ttgagtcaac gccactgtgc atgaagctcg taattgttat
     3061 ccgctcacaa ttaagggcga cacaaaattt attctaaatg ataataaata ctgataacat
     3121 cttatagttt gtattatatt ttgtattatc gttgacatgt ataattttga tatcaaaaac
     3181 tgattttccc tttattattt tcgagattta ttttcttaat tctctttaac aaactagaaa
     3241 tattgtatat acaaaaaatc ataaataata gatgaatagt ttaattatag gtgttcatca
     3301 atcgaaaaag caacgtatct tatttaaagt gcgttgcttt tttctcattt ataaggttaa
     3361 ataattctca tatatcaagc aaagtgacag gcgcccttaa atattctgac aaatgctctt
     3421 tccctaaact ccccccataa aaaaacccgc cgaagcgggt ttttacgtta tttgcggatt
     3481 aacgattact cgttatcaga accgcccagg gggcccgagc ttaagactgg ccgtcgtttt
     3541 acaacacaga aagagtttgt agaaacgcaa aaaggccatc cgtcaggggc cttctgctta
     3601 gtttgatgcc tggcagttcc ctactctcgc cttccgcttc ctcgctcact gactcgctgc
     3661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4201 gcggtgctac agagttcttg aagtggtggg ctaactacgg ctacactaga agaacagtat
     4261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4321 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4381 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4441 ggaacgacgc gcgcgtaact cacgttaagg gattttggtc atgagcaagg ggtgttatga
     4501 gccatattca ggtataaatg ggctcgcgat aatgttcaga attggttaat tggttgtaac
     4561 actgacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4621 ataaccctga taaatgcttc aataatattg aaaaaggaag aatatgagta ttcaacattt
     4681 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4741 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4801 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4861 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4921 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4981 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5041 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5101 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5161 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag cgatggcaac
     5221 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5281 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5341 ctggtttatt gctgataaat ccggagccgg tgagcgtggt tctcgcggta tcatcgcagc
     5401 gctggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5461 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5521 gtaatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag
     5581 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     5641 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg
     5701 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     5761 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgta
     5821 gcacgtgtca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct
     5881 cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg
     5941 atgtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga
     6001 cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc
     6061 acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca
     6121 gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt
     6181 cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct
     6241 tgccgatgtc gagcccgacg cgcgtgag
//
