LOCUS       sequence-368610-        6538 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 6538)
  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 6538)
  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-LAIR1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..115
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(134..181)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(229..558)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      243..378
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(297..316)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     543..563
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    681..792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     781..798
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(938..955)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(956..1000)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2249..2268)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2281..2299)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2282..2299)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2509..2533)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2530..2733)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2559..2579)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2734..3113
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3280..3296
                     /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(3752..3769)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3752..3768)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3867..3884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4038..4626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4118..4137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4712..4731)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4737..4756)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4798..4889
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4890..4958,4959..5750)
                     /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(5108..5127)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5803..5924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5814..5833)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5868..5887
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6054..6538,1..115))
                     /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(6137..6157)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
       61 ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggttt
      121 agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
      181 cacgtgctga tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc
      241 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc
      301 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
      361 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
      421 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
      481 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      541 acaccctaac tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc
      601 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
      661 gagatagggt tgagtgttgt gatgcaattt cctcatttta ttaggaaagg acagtgggag
      721 tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac
      781 tagaaggcac agtcgaggct gattttctct atctatttat tttactactc aagactatct
      841 tgcaaagtag aagaaccctg tctgcagtat gacgaccgtg tatacccgat tggcttaccg
      901 ggtaacgcgt tgtgccagtg actacaggcc ggccttagtg gtgatggtgg tggtgttcat
      961 gccactcaat cttctgagcc tcgaaaatat cattaagacc gctgccgccg gggctcaggc
     1021 tcagggactt ctgggtgtag tggttgtgca gggcctcgtg catcacgctg cagctgaaca
     1081 cgttgccctg ctgccacctg gacttgtcca cggtcagctt gctgtacagg aagaagctgc
     1141 cgtcgctgtc cagcactggg ggggtggtct tgtagttgtt ctcgggctgg ccgttgctct
     1201 cccactccac ggcgatatcg ctggggtaga agcccttcac cagacaggtc agggacacct
     1261 ggttcttggt catctcctcc cggctggggg gcagggtgta cacctggggc tcccgtggct
     1321 ggcccttggc cttgctgatt gtcttttcga ttggggcagc cagggccttg ttggagactt
     1381 tgcacttgta ttctttgccg ttcagccagt cctggtgcag cacggtcagc acggacacca
     1441 ccctgtaggt gctgttgtac tgctcctctc tgggcttggt cttggcgttg tgcacctcca
     1501 cgccgtccac gtaccagttg aacttcacct ctgggtcctc gtgggacacg gccaccacca
     1561 cgcaggtcac ctcgggggtc ctgctgatca tcagggtgtc cttgggcttg ggggggaaca
     1621 ggaacacgga aggccctccc agcagttctg gagctgggca gggggggcag gtgtgggtct
     1681 tgtcgccgcc cggcgcgcca tacagatgtt cggccttcag tccctgagag gcaggggcat
     1741 gctcattgtg agaattgtcg ctgggcctct gtgtaggtcc ggcagaagat ccaggctctg
     1801 tatcagggct gtcagggccg ccagatgtct ctttgaccag cagttccagg tagtcgctct
     1861 gctcggacca cttaggaggc ttgtagtaga tacaccggta aggtccggcg ttgccctcgg
     1921 acacgctgtc gattctgaat ctggcctcag actcagatgg agaggcctgg gacacgtcct
     1981 cggtgtcgtt gtaggtgctt ctgctctctc tttccagccg gaaggtctgc acgcccacag
     2041 gtcctctaca cacgaaggtc acgtggctgc ccagaggaat cactgttcca ggctcggcgc
     2101 tgatgctagg tctaggcaga tcttcttctt gggcggccgc agcgaacacc aggcagaaga
     2161 tgtagctcag ggcaatgatg gtcttcatgg tggcgctagc aggcgcgttt tcgatagaac
     2221 cgaggtgcag ttggacctgg gagtggacac ctgtggagag aaaggcaaag tggatgttat
     2281 tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc
     2341 gcgttctaac gaagccaagg gggtgggcct atagactcta taggcggtac ttacgtcact
     2401 cttggcacgg ggaatccgcg ttccaatgca ccgttcccgg ccgcggaggc tggatcggtc
     2461 ccggtgtctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga tctgacggtt
     2521 cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     2581 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2641 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     2701 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     2761 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     2821 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     2881 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     2941 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     3001 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     3061 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gccaacatgg
     3121 cggtcatatt ggacatgagc caatataaat gtacatatta tgatatagat acaacgtatg
     3181 caatggccaa tagccaatat tgatttatgc tatataacca atgaataata tggctaatgg
     3241 ccaatattga gtcaacgcca ctgtgcatga agctcgtaat tgttatccgc tcacaattaa
     3301 gggcgacaca aaatttattc taaatgataa taaatactga taacatctta tagtttgtat
     3361 tatattttgt attatcgttg acatgtataa ttttgatatc aaaaactgat tttcccttta
     3421 ttattttcga gatttatttt cttaattctc tttaacaaac tagaaatatt gtatatacaa
     3481 aaaatcataa ataatagatg aatagtttaa ttataggtgt tcatcaatcg aaaaagcaac
     3541 gtatcttatt taaagtgcgt tgcttttttc tcatttataa ggttaaataa ttctcatata
     3601 tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat gctctttccc taaactcccc
     3661 ccataaaaaa acccgccgaa gcgggttttt acgttatttg cggattaacg attactcgtt
     3721 atcagaaccg cccagggggc ccgagcttaa gactggccgt cgttttacaa cacagaaaga
     3781 gtttgtagaa acgcaaaaag gccatccgtc aggggccttc tgcttagttt gatgcctggc
     3841 agttccctac tctcgccttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3901 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3961 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4021 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4081 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4141 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4201 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4261 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4321 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4381 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4441 ttcttgaagt ggtgggctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4501 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4561 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4621 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgcgc
     4681 gtaactcacg ttaagggatt ttggtcatga gcaaggggtg ttatgagcca tattcaggta
     4741 taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg acccctattt
     4801 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4861 tgcttcaata atattgaaaa aggaagaata tgagtattca acatttccgt gtcgccctta
     4921 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4981 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5041 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5101 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5161 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5221 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5281 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5341 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5401 taccaaacga cgagcgtgac accacgatgc ctgtagcgat ggcaacaacg ttgcgcaaac
     5461 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5521 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5581 ataaatccgg agccggtgag cgtggttctc gcggtatcat cgcagcgctg gggccagatg
     5641 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5701 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa tgaccatgat
     5761 tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt
     5821 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     5881 ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac
     5941 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     6001 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtagcac gtgtcaggca
     6061 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     6121 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggatgt ctccaggaag
     6181 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     6241 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     6301 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     6361 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     6421 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     6481 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccg
//
