LOCUS       sequence-368850-        6493 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 6493)
  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 6493)
  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-MFAP3-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..120
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(139..186)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(234..563)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      248..383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(302..321)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     548..568
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    686..797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     786..803
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(943..960)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(961..1005)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2209..2228)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2241..2259)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2242..2259)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2469..2493)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2490..2693)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2519..2539)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2694..3073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3240..3256
                     /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(3712..3729)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3712..3728)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3827..3844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3998..4586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4078..4097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4672..4691)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4697..4716)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4758..4849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4850..4918,4919..5710)
                     /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(5068..5087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5763..5884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5774..5793)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5828..5847
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6014..6493,1..120))
                     /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(6097..6117)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
       61 tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
      121 ggtttagttc ctcaccttgt cgtattatac tatgccgata tactatgccg atgattaatt
      181 gtcaacacgt gctgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa
      241 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg
      301 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
      361 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      421 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      481 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      541 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
      601 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      661 agaccgagat agggttgagt gttgtgatgc aatttcctca ttttattagg aaaggacagt
      721 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
      781 gcaactagaa ggcacagtcg aggctgattt tctctatcta tttattttac tactcaagac
      841 tatcttgcaa agtagaagaa ccctgtctgc agtatgacga ccgtgtatac ccgattggct
      901 taccgggtaa cgcgttgtgc cagtgactac aggccggcct tagtggtgat ggtggtggtg
      961 ttcatgccac tcaatcttct gagcctcgaa aatatcatta agaccgctgc cgccggggct
     1021 caggctcagg gacttctggg tgtagtggtt gtgcagggcc tcgtgcatca cgctgcagct
     1081 gaacacgttg ccctgctgcc acctggactt gtccacggtc agcttgctgt acaggaagaa
     1141 gctgccgtcg ctgtccagca ctgggggggt ggtcttgtag ttgttctcgg gctggccgtt
     1201 gctctcccac tccacggcga tatcgctggg gtagaagccc ttcaccagac aggtcaggga
     1261 cacctggttc ttggtcatct cctcccggct ggggggcagg gtgtacacct ggggctcccg
     1321 tggctggccc ttggccttgc tgattgtctt ttcgattggg gcagccaggg ccttgttgga
     1381 gactttgcac ttgtattctt tgccgttcag ccagtcctgg tgcagcacgg tcagcacgga
     1441 caccaccctg taggtgctgt tgtactgctc ctctctgggc ttggtcttgg cgttgtgcac
     1501 ctccacgccg tccacgtacc agttgaactt cacctctggg tcctcgtggg acacggccac
     1561 caccacgcag gtcacctcgg gggtcctgct gatcatcagg gtgtccttgg gcttgggggg
     1621 gaacaggaac acggaaggcc ctcccagcag ttctggagct gggcaggggg ggcaggtgtg
     1681 ggtcttgtcg ccgcccggcg cgcccatgtc gccgctggtg aagatcactc tcagggtcac
     1741 gctgtagctg gctctgatgg ggcttgtcac gaaacaggtg tacaggcctc tgtcgtcgaa
     1801 ggccacgttg gtgatgttca ggaagttgtc ggacaccagc cattttccgc ctctggatct
     1861 gccatccagc tgctggccct tagaattgtg ccagtgcacg tcctcgtagt ggctggctgt
     1921 cagcagacac tcgatgctga cgctggtgcc ctctttggca atgatcacat cgtcgtcgct
     1981 gtggctgctg gcgctcagct caaaggagct gggaaagctg gcgttgtagc tgcttctgtt
     2041 ggcttccagg gacaccatct ggtcgaagtc cacatcttcc agcacaaaag cggcggccgc
     2101 agcgaacacc aggcagaaga tgtagctcag ggcaatgatg gtcttcatgg tggcgctagc
     2161 aggcgcgttt tcgatagaac cgaggtgcag ttggacctgg gagtggacac ctgtggagag
     2221 aaaggcaaag tggatgttat tctatagtgt cacctaaatc gtatgtgtat gatacataag
     2281 gttatgtatt aattgtagcc gcgttctaac gaagccaagg gggtgggcct atagactcta
     2341 taggcggtac ttacgtcact cttggcacgg ggaatccgcg ttccaatgca ccgttcccgg
     2401 ccgcggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg tggatggcgt
     2461 ctccaggcga tctgacggtt cactaaacga gctctgctta tatagacctc ccaccgtaca
     2521 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     2581 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     2641 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     2701 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     2761 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     2821 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     2881 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     2941 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     3001 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     3061 aataatcaat gccaacatgg cggtcatatt ggacatgagc caatataaat gtacatatta
     3121 tgatatagat acaacgtatg caatggccaa tagccaatat tgatttatgc tatataacca
     3181 atgaataata tggctaatgg ccaatattga gtcaacgcca ctgtgcatga agctcgtaat
     3241 tgttatccgc tcacaattaa gggcgacaca aaatttattc taaatgataa taaatactga
     3301 taacatctta tagtttgtat tatattttgt attatcgttg acatgtataa ttttgatatc
     3361 aaaaactgat tttcccttta ttattttcga gatttatttt cttaattctc tttaacaaac
     3421 tagaaatatt gtatatacaa aaaatcataa ataatagatg aatagtttaa ttataggtgt
     3481 tcatcaatcg aaaaagcaac gtatcttatt taaagtgcgt tgcttttttc tcatttataa
     3541 ggttaaataa ttctcatata tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat
     3601 gctctttccc taaactcccc ccataaaaaa acccgccgaa gcgggttttt acgttatttg
     3661 cggattaacg attactcgtt atcagaaccg cccagggggc ccgagcttaa gactggccgt
     3721 cgttttacaa cacagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggggccttc
     3781 tgcttagttt gatgcctggc agttccctac tctcgccttc cgcttcctcg ctcactgact
     3841 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4381 atgtaggcgg tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa
     4441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4621 ctcagtggaa cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gcaaggggtg
     4681 ttatgagcca tattcaggta taaatgggct cgcgataatg ttcagaattg gttaattggt
     4741 tgtaacactg acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4801 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagaata tgagtattca
     4861 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4921 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4981 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5041 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5101 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5161 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5221 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5281 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5341 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcgat
     5401 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5461 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5521 ggctggctgg tttattgctg ataaatccgg agccggtgag cgtggttctc gcggtatcat
     5581 cgcagcgctg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5641 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5701 gcattggtaa tgaccatgat tacgccaagc tctagctaga ggtcgacggt atacagacat
     5761 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     5821 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     5881 agttggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     5941 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     6001 ctcgtagcac gtgtcaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg
     6061 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
     6121 gcgcggatgt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
     6181 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca
     6241 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
     6301 cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc
     6361 ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc
     6421 acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc
     6481 gctcgatgtg gcg
//
