LOCUS       sequence-417867-        8839 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 8839)
  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 8839)
  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-MDGA1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4415..4459
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4460..4477
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4617..4634)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4623..4734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4852..4872)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4857..5186
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5037..5172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5099..5118
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5234..5281
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5300..5899
                     /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(5300..5319)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5796..5816
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6029..6150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6066..6085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6120..6139
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6203..6994,6995..7063))
                     /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     6826..6845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7064..7155)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7197..7216
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7222..7241
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7327..7915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7816..7835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8069..8086
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8184..8201
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8185..8201
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8657..8673
                     /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)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg cccaaggcgt ttacgctcct gctcaagctc agattgtgca
     1021 tgccggccag gcctgcgtgg tcaaagagga caatatcagc gagcgcgtgt acaccatcag
     1081 agaaggcgat accctgatgc tgcagtgcct ggtcacagga caccctagac ctcaagtgcg
     1141 gtggacaaaa acagccggca gcgccagcga caagttccaa gagacaagcg tgttcaacga
     1201 gacactgcgg atcgagcgga tcgccagaac acaaggcggc agatattact gcaaggccga
     1261 gaatggcgtg ggcgtgccag ccatcaagag catcagagtg gacgtgcagt acctggacga
     1321 gcctatgctg acagtgcacc agacagtgtc cgacgtgcgg ggcaacttct accaagaaaa
     1381 gaccgtgttc ctgaggtgca ccgtgaacag caaccctcca gccagattca tctggaagag
     1441 aggcagcgac acactgagcc acagccagga taacggcgtg gacatctacg agcctctgta
     1501 cacacagggc gagacaaagg tgctgaagct gaagaatctg cggccccagg actacgccag
     1561 ctatacttgt caggtgtccg tgcggaatgt gtgcggcatc cctgataagg ccatcacctt
     1621 ccggctgacc aacacaacag cccctccagc tctgaagctg tccgtgaatg agacactggt
     1681 ggtcaacccc ggcgagaatg tgactgtgca atgtctgctg acaggcggcg atcctctgcc
     1741 tcagcttcaa tggtcacatg gccctggacc tttgcctctg ggagcacttg ctcaaggcgg
     1801 aacactgtct attcccagcg tgcaggccag agacagcggc tactataatt gcaccgccac
     1861 caacaacgtg ggcaaccccg ccaagaaaac cgtgaatctg ctcgtgcgga gcatgaagaa
     1921 cgccaccttc cagatcaccc cagacgtgat caaagagagc gagaacatcc agctcggcca
     1981 ggacctgaaa ctgagctgcc atgtggatgc cgtgcctcaa gagaaagtga cctaccagtg
     2041 gttcaagaac ggcaagcccg ccagaatgag caagagactg ctggtcacca gaaacgatcc
     2101 agagctgcct gccgtgacaa gcagcctgga actgatcgac ctgcacttca gcgactacgg
     2161 cacctacctg tgcatggcta gttttccagg cgctcccgtg cctgatctga gcgtggaagt
     2221 gaacatcagc agcgagacag tgcctcctac catcagcgtg ccaaaaggca gagccgtcgt
     2281 gacagttaga gagggctctc ctgccgaact gcagtgtgaa gtgcggggaa aacccagacc
     2341 tcctgtgctg tggtcccgcg tggacaaaga agctgctctg ctgccttctg gactgcccct
     2401 ggaagaaaca cccgatggca agctgagact ggaacgggtg tccagagaca tgagcggcac
     2461 atacagatgc cagaccgcca gatacaacgg cttcaacgtg cggcctagag aagcccaggt
     2521 gcagctgaat gtgcagttcc ctccagaggt ggaacctagc tctcaggatg tgcgacaagc
     2581 tctgggtaga cccgtgctgc tcagatgttc tctgctgaga ggctcccctc agagaattgc
     2641 ctctgccgtg tggcggttta agggacaact tctgcctcct cctcctgtgg tgcctgctgc
     2701 tgctgaagca cctgatcacg ctgaactgag gctggacgcc gtgaccagag atagcagcgg
     2761 cagctacgag tgcagcgtgt ccaatgatgt gggaagcgcc gcctgcctgt ttcaagtgtc
     2821 tgccaaggcc tactctcccg agttctactt cgacacccct aatcctacca gaagccacaa
     2881 gctgagcaag aactacagct acgtcctgca gtggacccag agggaacctg atgcagtcga
     2941 ccccgtgctg aactacagac tgtccatcag acagctgaac cagcacaacg ccgtggtcaa
     3001 ggctatccct gtgcggagag tggaaaaggg ccagctgctc gagtacatcc tgaccgatct
     3061 gagagtgccc cactcctacg aagtgcggct gaccccttac acaaccttcg gagctggcga
     3121 catggccagc cggatcatcc actacaccga gcctatcaac agccctaacc tgagcgacaa
     3181 tacctgccac ttcgaggacg agaagatctg tggctacacc caggatctga ccgacaactt
     3241 cgactggacc agacagaacg ccctgacaca gaaccccaag cggagcccta ataccggacc
     3301 tcctaccgat atctctggca cccctgaggg ctactacatg ttcatcgaaa ccagcagacc
     3361 cagagagctg ggcgatagag ctagactggt gtccccactg tacaacgcct ccgccaagtt
     3421 ctactgcgtg tccttcttct accatatgta cggcaagcac atcggcagcc tgaacctgct
     3481 tgtgcggtca agaaacaaag gcgccctgga tacacacgct tggagcctgt ctggcaacaa
     3541 gggcaatgtg tggcagcagg cccatgtgcc tatcagccca tctggcccct ttcagatcat
     3601 cttcgagggc gtcagaggcc ctggctatct gggagatatc gccatcgacg acgtgaccct
     3661 gaagaaaggc gagtgcccta gaaagcagac agaccccaac aaggtggtgg tcatgggcgc
     3721 gccgggcggc gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg
     3781 gccttccgtg ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc
     3841 cgaggtgacc tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg
     3901 gtacgtggac ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa
     3961 cagcacctac agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa
     4021 agaatacaag tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag
     4081 caaggccaag ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga
     4141 gatgaccaag aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat
     4201 cgccgtggag tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt
     4261 gctggacagc gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg
     4321 gcagcagggc aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac
     4381 ccagaagtcc ctgagcctga gccccggcgg cagcggtctt aatgatattt tcgaggctca
     4441 gaagattgag tggcatgaac accaccacca tcaccactaa ggccggcctg tagtcactgg
     4501 cacaacgcgt tacccggtaa gccaatcggg tatacacggt cgtcatactg cagacagggt
     4561 tcttctactt tgcaagatag tcttgagtag taaaataaat agatagagaa aatcagcctc
     4621 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4681 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcacaaca
     4741 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4861 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4921 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4981 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5041 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5101 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5161 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5221 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     5281 caaggtgagg aactaaacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5341 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5401 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5461 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5521 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5581 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5641 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5701 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5761 cggggtgccc gccttcctgg agacatccgc gccccgcaac ctccccttct acgagcggct
     5821 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5881 ccgcaagccc ggtgcctgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     5941 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6001 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6061 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6121 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6181 gagcttggcg taatcatggt cattaccaat gcttaatcag tgaggcacct atctcagcga
     6241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6301 gggagggctt accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg
     6361 ctccggattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6481 cgccagttaa tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct
     6541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7021 caaaaaaggg aataagggcg acacggaaat gttgaatact catattcttc ctttttcaat
     7081 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7141 agaaaaataa acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg
     7201 cgagcccatt tatacctgaa tatggctcat aacacccctt gctcatgacc aaaatccctt
     7261 aacgtgagtt acgcgcgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7321 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7381 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7441 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttagccca
     7501 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7561 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7621 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7681 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7741 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7801 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7861 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7921 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7981 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     8041 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaaggcga
     8101 gagtagggaa ctgccaggca tcaaactaag cagaaggccc ctgacggatg gcctttttgc
     8161 gtttctacaa actctttctg tgttgtaaaa cgacggccag tcttaagctc gggccccctg
     8221 ggcggttctg ataacgagta atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg
     8281 gtttttttat ggggggagtt tagggaaaga gcatttgtca gaatatttaa gggcgcctgt
     8341 cactttgctt gatatatgag aattatttaa ccttataaat gagaaaaaag caacgcactt
     8401 taaataagat acgttgcttt ttcgattgat gaacacctat aattaaacta ttcatctatt
     8461 atttatgatt ttttgtatat acaatatttc tagtttgtta aagagaatta agaaaataaa
     8521 tctcgaaaat aataaaggga aaatcagttt ttgatatcaa aattatacat gtcaacgata
     8581 atacaaaata taatacaaac tataagatgt tatcagtatt tattatcatt tagaataaat
     8641 tttgtgtcgc ccttaattgt gagcggataa caattacgag cttcatgcac agtggcgttg
     8701 actcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
     8761 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
     8821 ccaatatgac cgccatgtt
//
