LOCUS       sequence-368734-        6610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            Fc(DAPA)-Avi-6xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  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 6610)
  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-SIRPD-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..49)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     276..296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    414..525
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     514..531
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(671..688)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(689..733)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2054..2073)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2086..2104)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2087..2104)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2314..2338)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2335..2538)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2364..2384)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2539..2918
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3085..3101
                     /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(3557..3574)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3557..3573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3672..3689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3843..4431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3923..3942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4517..4536)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4542..4561)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4603..4694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4695..4763,4764..5555)
                     /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(4913..4932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5608..5729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5619..5638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5673..5692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5859..6458)
                     /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(5942..5962)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6439..6458
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6477..6524)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6572..6610,1..291))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6586..6610,1..111)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
       61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      301 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
      361 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgtgatgcaa
      421 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
      481 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgattttc
      541 tctatctatt tattttacta ctcaagacta tcttgcaaag tagaagaacc ctgtctgcag
      601 tatgacgacc gtgtataccc gattggctta ccgggtaacg cgttgtgcca gtgactacag
      661 gccggcctta gtggtgatgg tggtggtgtt catgccactc aatcttctga gcctcgaaaa
      721 tatcattaag accgctgccg ccggggctca ggctcaggga cttctgggtg tagtggttgt
      781 gcagggcctc gtgcatcacg ctgcagctga acacgttgcc ctgctgccac ctggacttgt
      841 ccacggtcag cttgctgtac aggaagaagc tgccgtcgct gtccagcact gggggggtgg
      901 tcttgtagtt gttctcgggc tggccgttgc tctcccactc cacggcgata tcgctggggt
      961 agaagccctt caccagacag gtcagggaca cctggttctt ggtcatctcc tcccggctgg
     1021 ggggcagggt gtacacctgg ggctcccgtg gctggccctt ggccttgctg attgtctttt
     1081 cgattggggc agccagggcc ttgttggaga ctttgcactt gtattctttg ccgttcagcc
     1141 agtcctggtg cagcacggtc agcacggaca ccaccctgta ggtgctgttg tactgctcct
     1201 ctctgggctt ggtcttggcg ttgtgcacct ccacgccgtc cacgtaccag ttgaacttca
     1261 cctctgggtc ctcgtgggac acggccacca ccacgcaggt cacctcgggg gtcctgctga
     1321 tcatcagggt gtccttgggc ttggggggga acaggaacac ggaaggccct cccagcagtt
     1381 ctggagctgg gcaggggggg caggtgtggg tcttgtcgcc gcccggcgcg cctttgctca
     1441 gcaggcctgt cagtcccagc agtctcagac agcagcaggg ctgcacgaag taattggtgc
     1501 tgttccgctc aggcagggca gacagacatg tgtgagcatc atggtgggct ctgcttccgg
     1561 ctctgccagc aggtctgttc ttaggaggtc tggggttctg ctctgtcacg aacacctggg
     1621 tgcctctgcc gctctggtac tctttgatgg ctcggccctt gataaacttc acgcagtagt
     1681 aggtgccggc atcggccaga gagatttctc tgatccgggt gctgaagtcg gtgttgccag
     1741 gcttggtggt atcgccgatc tctttcactc tggggaagtt gccctgcttg aagttgtaga
     1801 tcagcttccg gttggggcct gtgcctttga accacagcac ggggccatta ggcagtgtgt
     1861 tgggcacgct gcagctcagg atgatgctct cgccggtaga cacggtctgg ctcatctctg
     1921 tctgctgcac atgaaaggcg gccgcagcga acaccaggca gaagatgtag ctcagggcaa
     1981 tgatggtctt catggtggcg ctagcaggcg cgttttcgat agaaccgagg tgcagttgga
     2041 cctgggagtg gacacctgtg gagagaaagg caaagtggat gttattctat agtgtcacct
     2101 aaatcgtatg tgtatgatac ataaggttat gtattaattg tagccgcgtt ctaacgaagc
     2161 caagggggtg ggcctataga ctctataggc ggtacttacg tcactcttgg cacggggaat
     2221 ccgcgttcca atgcaccgtt cccggccgcg gaggctggat cggtcccggt gtcttctatg
     2281 gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct
     2341 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     2401 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     2461 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     2521 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     2581 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     2641 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     2701 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     2761 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     2821 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     2881 aattgattac tattaataac tagtcaataa tcaatgccaa catggcggtc atattggaca
     2941 tgagccaata taaatgtaca tattatgata tagatacaac gtatgcaatg gccaatagcc
     3001 aatattgatt tatgctatat aaccaatgaa taatatggct aatggccaat attgagtcaa
     3061 cgccactgtg catgaagctc gtaattgtta tccgctcaca attaagggcg acacaaaatt
     3121 tattctaaat gataataaat actgataaca tcttatagtt tgtattatat tttgtattat
     3181 cgttgacatg tataattttg atatcaaaaa ctgattttcc ctttattatt ttcgagattt
     3241 attttcttaa ttctctttaa caaactagaa atattgtata tacaaaaaat cataaataat
     3301 agatgaatag tttaattata ggtgttcatc aatcgaaaaa gcaacgtatc ttatttaaag
     3361 tgcgttgctt ttttctcatt tataaggtta aataattctc atatatcaag caaagtgaca
     3421 ggcgccctta aatattctga caaatgctct ttccctaaac tccccccata aaaaaacccg
     3481 ccgaagcggg tttttacgtt atttgcggat taacgattac tcgttatcag aaccgcccag
     3541 ggggcccgag cttaagactg gccgtcgttt tacaacacag aaagagtttg tagaaacgca
     3601 aaaaggccat ccgtcagggg ccttctgctt agtttgatgc ctggcagttc cctactctcg
     3661 ccttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3721 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3781 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3841 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3901 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3961 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4021 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4081 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4141 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4201 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4261 gctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4321 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4381 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4441 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag
     4501 ggattttggt catgagcaag gggtgttatg agccatattc aggtataaat gggctcgcga
     4561 taatgttcag aattggttaa ttggttgtaa cactgacccc tatttgttta tttttctaaa
     4621 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4681 gaaaaaggaa gaatatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4741 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4801 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4861 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4921 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4981 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5041 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5101 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5161 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5221 gtgacaccac gatgcctgta gcgatggcaa caacgttgcg caaactatta actggcgaac
     5281 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5341 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tccggagccg
     5401 gtgagcgtgg ttctcgcggt atcatcgcag cgctggggcc agatggtaag ccctcccgta
     5461 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5521 ctgagatagg tgcctcactg attaagcatt ggtaatgacc atgattacgc caagctctag
     5581 ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa
     5641 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5701 taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc
     5761 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     5821 tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc aggcaccggg cttgcgggtc
     5881 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     5941 cgctcgtaga aggggaggtt gcggggcgcg gatgtctcca ggaaggcggg caccccggcg
     6001 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     6061 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     6121 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     6181 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     6241 accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     6301 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc
     6361 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg
     6421 cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg tttagttcct caccttgtcg
     6481 tattatacta tgccgatata ctatgccgat gattaattgt caacacgtgc tgatcagatc
     6541 cgaaaatgga tatacaagct cccgggagct ttttgcaaaa gcctaggcct ccaaaaaagc
     6601 ctcctcacta
//
