LOCUS       sequence-420989-        6661 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 6661)
  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 6661)
  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-TREM1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..116
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    234..345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     334..351
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(491..508)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(509..553)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1925..1944)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1957..1975)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1958..1975)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2185..2209)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2206..2409)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2235..2255)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2410..2789
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2956..2972
                     /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(3428..3445)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3428..3444)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3543..3560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3714..4302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3794..3813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4388..4407)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4413..4432)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4474..4565
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4566..4634,4635..5426)
                     /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(4784..4803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5479..5600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5490..5509)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5544..5563
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5730..6329)
                     /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(5813..5833)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6310..6329
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6348..6395)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6443..6661,1..111))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6457..6592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6511..6530)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
       61 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      121 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
      181 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgtgatgcaa
      241 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
      301 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgattttc
      361 tctatctatt tattttacta ctcaagacta tcttgcaaag tagaagaacc ctgtctgcag
      421 tatgacgacc gtgtataccc gattggctta ccgggtaacg cgttgtgcca gtgactacag
      481 gccggcctta gtggtgatgg tggtggtgtt catgccactc aatcttctga gcctcgaaaa
      541 tatcattaag accgctgccg ccggggctca ggctcaggga cttctgggtg tagtggttgt
      601 gcagggcctc gtgcatcacg ctgcagctga acacgttgcc ctgctgccac ctggacttgt
      661 ccacggtcag cttgctgtac aggaagaagc tgccgtcgct gtccagcact gggggggtgg
      721 tcttgtagtt gttctcgggc tggccgttgc tctcccactc cacggcgata tcgctggggt
      781 agaagccctt caccagacag gtcagggaca cctggttctt ggtcatctcc tcccggctgg
      841 ggggcagggt gtacacctgg ggctcccgtg gctggccctt ggccttgctg attgtctttt
      901 cgattggggc agccagggcc ttgttggaga ctttgcactt gtattctttg ccgttcagcc
      961 agtcctggtg cagcacggtc agcacggaca ccaccctgta ggtgctgttg tactgctcct
     1021 ctctgggctt ggtcttggcg ttgtgcacct ccacgccgtc cacgtaccag ttgaacttca
     1081 cctctgggtc ctcgtgggac acggccacca ccacgcaggt cacctcgggg gtcctgctga
     1141 tcatcagggt gtccttgggc ttggggggga acaggaacac ggaaggccct cccagcagtt
     1201 ctggagctgg gcaggggggg caggtgtggg tcttgtcgcc gcccggcgcg ccattgaaca
     1261 cgggcaccct gatgatgtcg gtcacgttgg tcaggttgat ctcgctgtcg ggtgtagaca
     1321 catcggcggt gctcttaggt ggggcctgtg tcactgttct ggggcttgtg tacagagggc
     1381 acagagcctt ggtggtggta ggagggatct tgtacacgtt ctgggtgctg ttctcgttgc
     1441 tgccaggggt gccagaaaag cccttggtga ccaccagtct gatccggtcg aacagcatgt
     1501 ggggttcttt aggaggctgg tagatcacac actggtacag gccgctatcc tcgacctgca
     1561 gattgaccat ccgcactctc agcaggccgt gatcgtggta atcttccagg atgatgcggc
     1621 ccacttgcac agggtgagaa ttcttgctgg gcctctcggt gcaggccagt gtcttaggca
     1681 tctcgccatc tctgatgatc tgccaggcct tctgagagct ggcgaacttt tccagggtgt
     1741 agtcgcactt cacgtccagt gtctggccct ctttcagctc gtacttctct tctgtcagct
     1801 ttgttgcggc ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct
     1861 tcatggtggc gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt
     1921 ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat
     1981 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt
     2041 gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc
     2101 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
     2161 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     2221 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     2281 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     2341 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     2401 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     2461 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     2521 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     2581 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     2641 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     2701 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     2761 ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat
     2821 ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat
     2881 ttatgctata taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt
     2941 gcatgaagct cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa
     3001 tgataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     3061 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     3121 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     3181 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     3241 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     3301 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
     3361 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
     3421 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
     3481 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
     3541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
     4141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4321 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
     4381 tcatgagcaa ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca
     4441 gaattggtta attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa
     4501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4561 agaatatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4801 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5101 cgatgcctgt agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg
     5281 gttctcgcgg tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta
     5341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5401 gtgcctcact gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc
     5461 gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5521 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5581 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     5641 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     5701 gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag
     5761 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     5821 aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc
     5881 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     5941 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     6001 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     6061 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     6121 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     6181 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     6241 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     6301 aggcgcaccg tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc gtattatact
     6361 atgccgatat actatgccga tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg
     6421 atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     6481 acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta
     6541 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     6601 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     6661 c
//
